Details of a Researcher - TOKINO Takashi

Molecular mechanisms of suppression of Claudin-1 expression in oral cancer cell lines

丹下正一朗, 井戸川雅史, 川島秀器, 佐々木泰史, 時野隆至

日本分子生物学会年会プログラム・要旨集(Web) 46th 2023

Identification of novel prognosticator gene in malignant pancreatic cancer

Shoichiro Tange, Tomomi Hirano, Masashi Idogawa, Eishu Hirata, Issei Imoto, Takashi Tokino

CANCER SCIENCE ( WILEY ) 113 2022.02

Research paper, summary (international conference)

膵臓がんにおける新規予後予測遺伝子の機能解析

丹下正一朗, 平野朋美, 井戸川雅史, 平田英周, 井本逸勢, 時野隆至

日本癌学会総会記事 ( (一社)日本癌学会 ) 80回 [J15 - 6] 2021.09

症例特異的変異を用いた頭頸部扁平上皮癌におけるctDNAモニタリング

古後龍之介, 真子知美, 岩谷岳, 西塚哲, 佐々木泰史, 井戸川雅史, 時野隆至, 中川尚志

日本癌学会総会記事 ( (一社)日本癌学会 ) 80回 [J14 - 6] 2021.09

RAS野生型転移性大腸癌患者における循環腫瘍DNA中のRAS、BRAF、PIK3CA変異の同定と腫瘍組織の変異との比較

澤田武, 久保田英嗣, 中村慶史, 高橋直樹, 太田亮介, 井戸川雅史, 佐々木泰史, 時野隆至, 源利成, 片岡洋望

日本癌学会総会記事 ( (一社)日本癌学会 ) 80回 [P15 - 6] 2021.09

長鎖非コードRNA lncAC1は大腸癌の進行を促進する

小泉昌代, 井戸川雅史, 丹下正一朗, 時野隆至

日本癌学会総会記事 ( (一社)日本癌学会 ) 80回 [P14 - 5] 2021.09

カスタムパネルシーケンスを用いた日本人メラノーマのドライバー遺伝子の検出

肥田時征, 井戸川雅史, 堀本浩平, 加藤潤史, 佐藤さゆり, 藤岡茉生, 丹下正一朗, 時野隆至, 宇原久

日本皮膚科学会雑誌 ( (公社)日本皮膚科学会 ) 131 ( 5 ) 1363 - 1363 2021.05

日本人口腔扁平上皮癌における全exome解析

佐々木泰史, 中垣貴文, 中垣貴文, 荻和弘, 丹下正一朗, 井戸川雅史, 宮崎晃亘, 時野隆至

日本分子生物学会年会プログラム・要旨集(Web) 42nd 2019

Digital PCRを用いた食道癌患者における症例特異的血漿中遊離DNAモニタリング(Patient-specific circulating tumor DNA monitoring using digital PCR in esophageal squamous cell cancer patients)

岩谷岳, 遠藤史隆, 佐々木泰史, 八重樫瑞典, 佐藤慧, 佐々木教之, 秋山有史, 佐々木章, 増田万里, 時野隆至, 西塚哲

日本癌学会総会記事 ( 日本癌学会 ) 77回 930 - 930 2018.09

食道癌肉腫の遺伝子変異解析(Exome sequence analysis of carcinomatous and sarcomatous elements of an esophageal carcinosarcoma)

佐々木教之, 岩谷岳, 遠藤史隆, 鴻巣正史, 秋山有史, 佐々木泰史, 時野隆至, 西塚哲

日本癌学会総会記事 ( 日本癌学会 ) 77回 326 - 326 2018.09

新規p53標的遺伝子として同定されたp120カテニンファミリー蛋白の癌における役割

鈴木菜摘, 井戸川雅史, 大箸智子, 佐々木泰史, 時野隆至

日本分子生物学会年会プログラム・要旨集(Web) 41st 2018

胃がん発生に関与する長鎖non-coding RNAの同定

北嶋洋志, 丸山玲緒, 山本英一郎, 新沼猛, 甲斐正広, 時野隆至, 仲瀬裕志, 鈴木拓

生命科学系学会合同年次大会 ( 生命科学系学会合同年次大会運営事務局 ) 2017年度 [2P - 0828] 2017.12

慢性胃炎および胃癌に関する長鎖noncoding RNAの同定と機能解析

北嶋洋志, 丸山玲緒, 山本英一郎, 新沼猛, 青木敬則, 甲斐正広, 時野隆至, 今井浩三, 仲瀬裕志, 鈴木拓

日本癌学会総会記事 ( 日本癌学会 ) 76回 P - 3231 2017.09

慢性B型肝炎の核酸アナログ製剤治療後発がんに関与するmicroRNAの探索

若杉英樹, 鈴木拓, 佐々木基, 新沼猛, 山本英一郎, 甲斐正広, 高橋秀明, 伊東文生, 時野隆至, 仲瀬裕志

日本癌学会総会記事 ( 日本癌学会 ) 76回 P - 3310 2017.09

消化管間質腫瘍においてエピジェネティックに制御される長鎖noncoding RNAの探索

新沼猛, 北嶋洋志, 山本英一郎, 甲斐正広, 仲瀬裕志, 時野隆至, 今井浩三, 鈴木拓

日本癌学会総会記事 ( 日本癌学会 ) 76回 P - 2257 2017.09

新規大腸がん線維芽細胞関連遺伝子の同定

沼田有斗, 萬顕, 山本英一郎, 新沼猛, 北嶋洋志, 甲斐正広, 青木敬則, 若杉英樹, 時野隆至, 仲瀬裕志, 菅井有, 鈴木拓

日本癌学会総会記事 ( 日本癌学会 ) 76回 P - 2219 2017.09

大腸腫瘍の表面構造が反映する腫瘍内不均一性

山本英一郎, 山野泰穂, 青木敬則, 新沼猛, 甲斐正広, 佐々木泰史, 時野隆至, 菅井有, 仲瀬裕志, 鈴木拓

日本癌学会総会記事 ( 日本癌学会 ) 76回 P - 2228 2017.09

食道癌患者におけるCirculating tumor DNAを用いたLiquid biopsy systemの開発

岩谷岳, 遠藤史隆, 佐々木泰史, 八重樫瑞典, 千葉丈広, 秋山有史, 増田万里, 山田哲司, 時野隆至, 西塚哲

日本癌学会総会記事 ( 日本癌学会 ) 76回 J - 3064 2017.09

大腸がんにおける腫瘍血管内皮関連遺伝子の同定

萬顕, 山本英一郎, 沼田有斗, 新沼猛, 北嶋洋志, 甲斐正広, 青木敬則, 若杉英樹, 時野隆至, 中瀬裕志, 菅井有, 鈴木拓

日本癌学会総会記事 ( 日本癌学会 ) 76回 P - 2224 2017.09

DOWNREGULATION OF MIR-200B IS ASSOCIATED WITH CISPLATIN-RESISTANCE IN BLADDER CANCER CELLS

Tetsuya Shindo, Naotaka Nishiyama, Takeshi Niinuma, Hiroshi Kitajima, Masahiro Kai, Takashi Tokino, Nobuo Shinkai, Hiromu Suzuki, Naoya Masumori

JOURNAL OF UROLOGY ( ELSEVIER SCIENCE INC ) 197 ( 4 ) E568 - E569 2017.04

Research paper, summary (international conference)

多発性骨髄腫に対するDot1L阻害剤の抗腫瘍効果解明を目指したエピゲノム解析

石黒一也, 北嶋洋志, 新沼猛, 丸山玲緒, 甲斐正広, 池田博, 石田禎夫, 佐々木泰史, 時野隆至, 仲瀬裕志, 鈴木拓

エピゲノムはどこまで操れるようになったか第11回日本エピジェネティクス研究会年会プログラム集理研シンポジウム平成29年 2017

慢性胃炎および胃癌に関連する長鎖noncoding RNAの同定

北嶋洋志, 丸山玲緒, 山本英一郎, 新沼猛, 青木敬則, 原田拓, 甲斐正広, 仲瀬裕志, 時野隆至, 鈴木拓

日本癌学会総会記事 ( 日本癌学会 ) 75回 P - 1225 2016.10

大腸癌における腫瘍血管内皮関連遺伝子の同定

萬顕, 山本英一郎, 津矢田明泰, 沼田有斗, 甲斐正広, 新沼猛, 北嶋洋志, 青木敬則, 若杉秀樹, 時野隆至, 仲瀬裕志, 菅井有, 鈴木拓

日本癌学会総会記事 ( 日本癌学会 ) 75回 P - 1282 2016.10

Targeted semiconductor sequencing of 409 cancer-related genes for somatic mutations and copy number variations in multiple myeloma

Hiroshi Ikeda, Yasushi Sasaki, Kazuya Ishiguro, Tadao Ishida, Yuka Aoki, Takashi Tokino

CANCER RESEARCH ( AMER ASSOC CANCER RESEARCH ) 76 2016.07

Research paper, summary (international conference)

LIMA1は新規p53標的遺伝子であり癌細胞浸潤を抑制する

大箸智子, 井戸川雅史, 佐々木泰史, 時野隆至

日本分子生物学会年会プログラム・要旨集(Web) 39th 2016

長鎖非コードRNAのフロンティア生化学、分子生物学、医学からのアプローチ慢性胃炎から胃癌への発癌過程に関与しうる長鎖非コードRNAの網羅的探索と病的意義の解明

丸山玲緒, 北嶋洋志, 山本英一郎, 佐藤由梨, 粂川昂平, 新沼猛, 甲斐正広, 篠村恭久, 時野隆至, 鈴木拓

日本生化学会大会・日本分子生物学会年会合同大会講演要旨集 ( (公社)日本生化学会 ) 88回・38回 [2W15 - p 2015.12

慢性胃炎から胃癌への発癌過程に関与する長鎖非コードRNAの同定と機能解析の試み

佐藤由梨, 丸山玲緒, 北嶋洋志, 山本英一郎, 新沼猛, 粂川昂平, 甲斐正広, 能正勝彦, 井戸川雅史, 佐々木泰史, 篠村恭久, 時野隆至, 鈴木拓

日本癌学会総会記事 ( 日本癌学会 ) 74回 E - 1172 2015.10

大腸癌においてDNAメチル化により抑制されている長鎖非コードRNAの網羅的同定

石黒一也, 粂川昂平, 丸山玲緒, 山本英一郎, 津矢田明泰, 進藤哲哉, 新沼猛, 甲斐正広, 山野泰穂, 菅井有, 時野隆至, 篠村恭久, 鈴木拓

日本癌学会総会記事 ( 日本癌学会 ) 74回 E - 1165 2015.10

The effect of forced expression of k-ras mutation on gastrointestinal cancer cells and IGF-IR targeting therapy

Yasushi Adachi, Yasutaka Matsunaga, Yasushi Sasaki, Katsuhiko Nosho, Hiroyuki Yamamoto, Yoshiaki Arimura, Takao Endo, Yasuo Kato, Takashi Tokino, David P. Carbone, Yasuhisa Shinomura

CANCER RESEARCH ( AMER ASSOC CANCER RESEARCH ) 75 2015.08

Research paper, summary (international conference)

Molecular diagnostics of drug resistant multiple myeloma cases using targeted next generation sequencing

Hiroshi Ikeda, Yasushi Sasaki, Tetsuyuki Igarashi, Yuka Aoki, Toshiaki Hayashi, Tadao Ishida, Takashi Tokino, Yasuhisa Sinomura

CANCER RESEARCH ( AMER ASSOC CANCER RESEARCH ) 75 2015.08

Research paper, summary (international conference)

Interaction between monocytes and bone marrow microenvironment in pathogenesis of multiple myeloma

Hiroshi Ikeda, Yuka Aoki, Toshiaki Hyayashi, Yumiko Maruyama, Tadao Ishida, Takashi Tokino, Yasuhisa Shinomura, Yasushi Sasaki

CANCER RESEARCH ( AMER ASSOC CANCER RESEARCH ) 74 ( 19 ) 2014.10

Research paper, summary (international conference)

Close Up実験法 Series252 Clinical Genomicsを目指した半導体NGSによるがんゲノム解析

大腸癌においてDNAメチル化により抑制されているlincRNAの網羅的同定(Genome-wide identification of lincRNAs epigenetically silenced by DNA methylation in colon cancer)

五十嵐哲祥, 粂川昂平, 丸山玲緒, 山本英一郎, 津矢田明泰, 鈴木亮, 新沼猛, 甲斐正広, 山野泰穂, 菅井有, 時野隆至, 篠村恭久, 鈴木拓

日本癌学会総会記事 ( 日本癌学会 ) 73回 P - 1126 2014.09

慢性胃炎から胃癌への発癌過程に関与する長鎖ncRNAの網羅的探索と機能解析の試み(Systematic identification of long non-coding RNAs involved in gastritis and gastric tumorigenesis)

丸山玲緒, 北嶋洋志, 山本英一郎, 新沼猛, 萬顕, 粂川昂平, 津矢田明泰, 鈴木亮, 甲斐正広, 能正勝彦, 時野隆至, 篠村恭久, 鈴木拓

日本癌学会総会記事 ( 日本癌学会 ) 73回 E - 2096 2014.09

丸喜明, 佐々木泰史, 中村友紀, 横井左奈, 大平美紀, 時野隆至, 永瀬浩喜

実験医学 32 ( 11 ) 1781 - 1788 2014.07

消化器癌の発生や進展に関与する長鎖ncRNAの量的・質的異常の探索と臨床応用への試み

次世代半導体シーケンサーを用いた口腔扁平上皮癌におけるがん関連遺伝子の変異解析

中垣貴文, 佐々木泰史, 井戸川雅史, 竹田康佑, 田村みゆき, 大箸智子, 荻和弘, 平塚博義, 時野隆至

Personalized Medicine Universe. Japanese Edition ( (一社)国際個別化医療学会 ) 3 ( Suppl.1 ) 51 - 51 2014.06

p53の直接転写標的AKR1B10は大腸癌で発現抑制されp53誘導アポトーシスを制御する

大箸智子, 井戸川雅史, 佐々木泰史, 時野隆至

Personalized Medicine Universe. Japanese Edition ( (一社)国際個別化医療学会 ) 3 ( Suppl.1 ) 50 - 50 2014.06

丸山玲緒, 山本英一郎, 粂川昂平, 津矢田明泰, 鈴木亮, 芦田仁己, 甲斐正広, 佐藤亜紀子, 新沼猛, 山野泰穂, 菅井有, 篠村恭久, 時野隆至, 鈴木拓

日本分子腫瘍マーカー研究会誌 ( 日本分子腫瘍マーカー研究会 ) 29 71 - 72 2014.04

DOI CiNii

Ion PGMシーケンサーを用いたがん関連遺伝子の網羅的変異解析

小橋建太, 佐々木泰史, 中垣貴文, 竹田康佑, 田村みゆき, 大箸智子, 井戸川雅史, 時野隆至

日本分子生物学会年会プログラム・要旨集(Web) 37th 2014

p53標的遺伝子AKR1B10はp53誘導性アポトーシスを増強することで大腸発癌を抑制する

大箸智子, 井戸川雅史, 佐々木泰史, 時野隆至

日本分子生物学会年会プログラム・要旨集(Web) 37th 2014

ゲノム網羅的p53結合領域解析とマイクロアレイを組み合わせたp53標的長鎖非コードRNA(lincRNA)の同定と解析

井戸川雅史, 大箸智子, 佐々木泰史, 時野隆至

日本分子生物学会年会プログラム・要旨集(Web) 37th 2014

Genomic vulnerability to LINE-1 hypomethylation is a potential determinant of the clinicogenetic features of multiple myeloma (vol 4, 101, 2012)

消化器癌において重要な役割を果たす長鎖ncRNAの網羅的同定の試み(Global identification of novel long non-coding RNAs potentially involved in gastrointestinal cancer)

丸山玲緒, 山本英一郎, 粂川昂平, 津矢田明泰, 鈴木亮, 芦田仁己, 佐藤亜紀子, 甲斐正広, 山野泰穂, 菅井有, 篠村恭久, 時野隆至, 鈴木拓

日本癌学会総会記事 ( 日本癌学会 ) 72回 124 - 124 2013.10

多発性骨髄腫におけるLINE-1異常低メチル化と臨床遺伝子学的特徴の相関(LINE-1 hypomethylation is a potential determinant of the clinicogenetic features of multiple myeloma)

野島正寛, 青木由佳, 安井寛, 丸山玲緒, 山本英一郎, 麻奥英毅, 時野隆至, 長村文孝, 石田禎夫, 今井浩三, 篠村恭久, 鈴木拓

日本癌学会総会記事 ( 日本癌学会 ) 72回 305 - 305 2013.10

Yuka Aoki, Masanori Nojima, Hiromu Suzuki, Hiroshi Yasui, Reo Maruyama, Eiichiro Yamamoto, Masami Ashida, Mitsuhiro Itagaki, Hideki Asaoku, Hiroshi Ikeda, Toshiaki Hayashi, Kohzoh Imai, Mitsuru Mori, Takashi Tokino, Tadao Ishida, Minoru Toyota, Yasuhisa Shinomura

GENOME MEDICINE ( BIOMED CENTRAL LTD ) 5 ( 10 ) 2013.10

消化管内視鏡の生検検体におけるヒストン修飾の網羅的な解析と新規分子マーカー同定の試み

消化器癌の発生や進展に関与する長鎖ncRNAの量的・質的異常の探索と臨床応用への試み

丸山玲緒, 山本英一郎, 粂川昂平, 津矢田明泰, 鈴木亮, 芦田仁己, 甲斐正広, 佐藤亜紀子, 新沼猛, 山野泰穂, 菅井有, 篠村恭久, 時野隆至, 鈴木拓

日本分子腫瘍マーカー研究会プログラム・講演抄録 ( 日本分子腫瘍マーカー研究会 ) 33回 100 - 101 2013.09

丸山玲緒, 山本英一郎, 粂川昂平, 井戸川雅史, 野島正寛, 甲斐正広, 佐藤亜紀子, 佐々木泰史, 山野泰穂, 菅井有, 篠村恭久, 時野隆至, 鈴木拓

日本分子腫瘍マーカー研究会誌 ( 日本分子腫瘍マーカー研究会 ) 28 16 - 17 2013.03

ゲノム網羅的p53結合領域解析によるp53ファミリーの転写標的となる大型遺伝子介在性非コードRNA(lincRNA)の同定と機能解析

井戸川雅史, 大箸智子, 佐々木泰史, 時野隆至

日本分子生物学会年会プログラム・要旨集(Web) 36th 2013

p53によるXPO1の抑制機構とその意義

佐々木泰史, 佐々木泰史, 小橋健太, 鈴木信太郎, 田村みゆき, 竹田康祐, 井戸川雅史, 井戸川雅史, 篠村恭久, 時野隆至

日本分子生物学会年会プログラム・要旨集(Web) 36th 2013

腫瘍マーカーとしてのエピジェネティクス研究の展望消化管内視鏡の生検検体におけるヒストン修飾の網羅的な解析と新規分子マーカー同定の試み

丸山玲緒, 山本英一郎, 粂川昂平, 井戸川雅史, 野島正寛, 甲斐正広, 佐藤亜紀子, 佐々木泰史, 山野泰穂, 菅井有, 篠村恭久, 時野隆至, 鈴木拓

日本分子腫瘍マーカー研究会プログラム・講演抄録 ( 日本分子腫瘍マーカー研究会 ) 32回 33 - 34 2012.09

大腸癌においてエピジェネティックに抑制されている長鎖ncRNAの網羅的同定の試み(Global identification of novel long non-coding RNAs silenced by epigenetic mechanism in colon cancer)

丸山玲緒, 粂川昂平, 山本英一郎, 井戸川雅史, 野島正寛, 甲斐正広, 能正勝彦, 佐々木泰史, 山野泰穂, 菅井有, 篠村恭久, 時野隆至, 鈴木拓

日本癌学会総会記事 ( 日本癌学会 ) 71回 381 - 381 2012.08

microRNA-196aとHOTAIRの過剰発現は消化管間質腫瘍の悪性度と相関する(Upregulation of miR-196a and HOTAIR drive malignant character in gastrointestinal stromal tumors)

鈴木拓, 新沼猛, 野島正寛, 丸山玲緒, 山本英一郎, 甲斐正広, 能正勝彦, 山本博幸, 時野隆至, 今井浩三, 篠村恭久

日本癌学会総会記事 ( 日本癌学会 ) 71回 419 - 419 2012.08

大腸腫瘍におけるニューロテンシン受容体1型遺伝子のメチル化と臨床的意義(Epigenetic silencing of neurotensin receptor type 1 (NTSR1) in colorectal tumor and its clinical implications)

山本英一郎, 鈴木拓, 神前正幸, 丸山玲緒, 山野泰穂, 澤田武, 能正勝彦, 山本博幸, 甲斐正広, 時野隆至, 菅井有, 今井浩三, 篠村恭久

日本癌学会総会記事 ( 日本癌学会 ) 71回 302 - 303 2012.08

p53ファミリーを用いた人工ハイブリッド遺伝子の作成とアポトーシス誘導活性

佐々木泰史, 佐々木泰史, 大島雄一郎, 小山良太, 田村みゆき, 井戸川雅史, 井戸川雅史, 篠村恭久, 時野隆至

日本分子生物学会年会プログラム・要旨集(Web) 35th 2012

がんの浸潤・遊走能に関わるp53ファミリーの新規標的FOXF1の同定と機能解析

田村みゆき, 佐々木泰史, 大箸智子, 井戸川雅史, 時野隆至

日本分子生物学会年会プログラム・要旨集(Web) 35th 2012

ゲノム網羅的shRNAスクリーニングによるp53変異癌細胞でのアポトーシス誘導

大箸智子, 井戸川雅史, 杉坂淳, 田村みゆき, 佐々木泰史, 時野隆至

日本分子生物学会年会プログラム・要旨集(Web) 35th 2012

Overexpression of Aurora A by loss of CHFR gene expression increases the growth and survival of HTLV-1-infected T cells through enhanced NF-kappaB activity (Retraction of vol 124, pg 2607, 2009)

M. Tomita, M. Toyota, C. Ishikawa, T. Nakazato, T. Okudaira, T. Matsuda, J. N. Uchihara, N. Taira, K. Ohshiro, M. Senba, Y. Tanaka, K. Ohshima, H. Saya, T. Tokino, N. Mori

INTERNATIONAL JOURNAL OF CANCER ( WILEY-BLACKWELL ) 129 ( 11 ) 2763 - 2763 2011.12

Other

大腸洗浄液を用いた新しい大腸癌の診断方法(Epigenetic alteration of DNA in mucosal wash fluid predicts invasiveness of colorectal tumors)

神前正幸, 山本英一郎, 鈴木拓, 山野泰穂, 甲斐正広, 菅井有, 時野隆至, 今井浩三, 篠村恭久, 豊田実

日本癌学会総会記事 ( 日本癌学会 ) 70回 135 - 135 2011.09

大腸癌のエピゲノム解析からアプローチする癌関連miRNAの探索(Genome-wide profiling of chromatin signatures reveals epigenetic regulation of microRNA genes in colorectal cancer)

鈴木拓, 山本英一郎, 野島正寛, 丸山玲緒, 高丸博之, 甲斐正広, 時野隆至, 今井浩三, 豊田実, 篠村恭久

日本癌学会総会記事 ( 日本癌学会 ) 70回 88 - 89 2011.09

異なる前癌病変に由来したCIMP大腸癌の亜型(Different subgroups of colorectal cancer with CIMP arise from genetically and epigenetically distinct early lesions)

山本英一郎, 鈴木拓, 山野泰穂, 神前正幸, 澤田武, 丸山玲緒, 甲斐正広, 菅井有, 時野隆至, 今井浩三, 篠村恭久, 豊田実

日本癌学会総会記事 ( 日本癌学会 ) 70回 46 - 46 2011.09

がんのエピジェネティクス基礎研究と臨床応用の進歩エピゲノム解析による造血器腫瘍の新たな分子標的の同定(Exploration of novel molecular targets for the therapy against hematological malignancies through epigenome analysis)

豊田実, 野島正寛, 鈴木拓, 丸山玲緒, 山本英一郎, 甲斐正広, 時野隆至, 篠村恭久, 今井浩三

日本癌学会総会記事 ( 日本癌学会 ) 70回 254 - 255 2011.09

乳癌においてDNAメチル化により不活化される遺伝子のゲノム網羅的解析

西川紀子, 豊田実, 鈴木拓, 藤兼智子, 九冨五郎, 亀嶋秀和, 鈴木やすよ, 大村東生, 時野隆至, 平田公一

日本外科学会雑誌 ( (一社)日本外科学会 ) 112 ( 臨増1-2 ) 393 - 393 2011.05

A comprehensive analysis to screen for epigenetically silenced miRNA genes in colorectal cancer

Hiromu Suzuki, Eiichiro Yamamoto, Masanori Nojima, Takashi Shimizu, Takashi Tokino, Yasuhisa Shinomura, Minoru Toyota

CANCER RESEARCH ( AMER ASSOC CANCER RESEARCH ) 71 2011.04

Research paper, summary (international conference)

Molecular mechanisms of anti-tumor activity of RAS association domain protein, RASSF2

豊田実, 丸山玲緒, 時野隆至

生体の科学 ( 金原一郎記念医学医療振興財団 ) 61 ( 6 ) 590 - 593 2010.11

拡大内視鏡によるCIMP陽性大腸癌の前がん病変の診断(Identification of precursor lesions of colorectal cancers with CIMP by molecular-magnifying colonoscopy)

神前正幸, 山本英一郎, 鈴木拓, 山野泰穂, 甲斐正広, 菅井有, 時野隆至, 今井浩三, 篠村恭久, 豊田実

日本癌学会総会記事 ( 日本癌学会 ) 69回 340 - 340 2010.08

GISTにおけるLINE-1低メチル化と悪性度との相関(A novel correlation between LINE-1 hypomethylation and the malignancy of gastrointestinal stromal tumors)

新沼猛, 鈴木拓, 野島正寛, 五十嵐伸一, 山本英一郎, 高丸博之, 田沼徳真, 山本博幸, 時野隆至, 今井浩三, 豊田実, 篠村恭久

日本癌学会総会記事 ( 日本癌学会 ) 69回 341 - 341 2010.08

染色体不安定性に関連した大腸腺腫内の異常メチル化(Epigenetic field defect in colorectal adenoma associated with chromosomal instability)

澤田武, 山本英一郎, 鈴木拓, 山野泰穂, 神前正幸, 甲斐正広, 菅井有, 時野隆至, 今井浩三, 篠村恭久, 伊東文生, 豊田実

日本癌学会総会記事 ( 日本癌学会 ) 69回 44 - 44 2010.08

CIMP大腸癌の起源(The origin of colorectal cancer with CpG island methylator phenotype)

山本英一郎, 鈴木拓, 山野泰穂, 神前正幸, 甲斐正広, 菅井有, 時野隆至, 今井浩三, 篠村恭久, 豊田実

日本癌学会総会記事 ( 日本癌学会 ) 69回 43 - 43 2010.08

NEGATIVE REGULATION OF HEPATOMA-DERIVED GROWTH FACTOR BY P53

Y. Sasaki, H. Negishi, R. Koyama, M. Kusano, H. Suzuki, M. Fujita, M. Toyota, T. Tokino

TUMOR BIOLOGY ( SPRINGER ) 31 S74 - S75 2010.08

Research paper, summary (international conference)

乳癌においてDNAメチル化により不活化される遺伝子のゲノム網羅的解析

西川紀子, 豊田実, 鈴木拓, 里見蕗乃, 藤兼智子, 九冨五郎, 亀島秀和, 鈴木やすよ, 大村東生, 時野隆至, 平田公一

日本乳癌学会総会プログラム抄録集 ( (一社)日本乳癌学会 ) 18回 354 - 354 2010.05

Epigenetic silencing of microRNA-34b/c in human gastric cancer

Hiromu Suzuki, Eiichiro Yamamoto, Masanori Nojima, Takashi Tokino, Kohzoh Lnnai, Minoru Toyota, Yasuhisa Shinomura

CANCER RESEARCH ( AMER ASSOC CANCER RESEARCH ) 70 2010.04

Research paper, summary (international conference)

CHFR, a potential tumor suppressor, downregulates interleukin-8 via inhibition of NF-kappa B

Takashi Tokino, Lisa Kashima, Minoru Toyota, Yasushi Sasaki

CANCER RESEARCH ( AMER ASSOC CANCER RESEARCH ) 70 2010.04

Research paper, summary (international conference)

DNA障害におけるPARP-1とKu70によるWnt/β-cateninシグナルの拮抗的制御

井戸川雅史, 山田哲司, 篠村恭久, 広橋説雄, 今井浩三, 時野隆至

日本分子腫瘍マーカー研究会誌 ( 日本分子腫瘍マーカー研究会 ) 25 44 - 46 2010.02

DNA障害におけるPARP-1とKu70によるWnt/β-cateninシグナルの拮抗的制御

井戸川雅史, 山田哲司, 篠村恭久, 広橋説雄, 今井浩三, 時野隆至

日本分子腫瘍マーカー研究会プログラム・講演抄録 ( 日本分子腫瘍マーカー研究会 ) 29回 61 - 63 2009.09

胃癌、大腸癌におけるIRFファミリーのエピジェネティックな不活化(Epigenetic inactivation of interferon regulatory factors (IRFs) in colorectal and gastric cancer)

山下真幸, 豊田実, 鈴木拓, 野島正寛, 渡邊嘉行, 明石浩史, 山本英一郎, 佐々木泰史, 菅井有, 篠村恭久, 今井浩三, 時野隆至, 伊東文生

日本癌学会総会記事 ( 日本癌学会 ) 68回 174 - 174 2009.08

大腸癌の多段階腫瘍形成における異常DNAメチル化の役割(The role of aberrant DNA methylation in multistep carcinogenesis of colorectal cancer)

山本英一郎, 山野泰穂, 鈴木拓, 野島正寛, 丸山怜緒, 時野隆至, 今井浩三, 篠村恭久, 豊田実

日本癌学会総会記事 ( 日本癌学会 ) 68回 94 - 94 2009.08

脱メチル化による遺伝子上方制御の全ゲノムスクリーニングによる乳癌の後成的サイレンシング標的の同定(Genomic Screening for Genes Upregulated by Demethylation Identified Targets of Epigenetic Silencing in Breast Cancer)

西川紀子, 豊田実, 鈴木拓, 藤兼智子, 野島正寛, 豊田睦美, 西舘敏彦, 佐々木泰, 平田公一, 時野隆至

日本癌学会総会記事 ( 日本癌学会 ) 68回 178 - 179 2009.08

IGFBP7はCpGアイランドメチル化因子表現型をもつ大腸癌において特異的にサイレンシングされるp53応答性遺伝子である(IGFBP7 is a p53 responsive gene specifically silenced in colorectal cancer with CpG island methylator phenotype)

丸山玲緒, 鈴木拓, 野島正寛, 山本英一郎, 五十嵐伸一, 高丸博之, 甲斐正広, 山本博幸, 伊東文生, 時野隆至, 今井浩三, 豊田実, 篠村恭久

日本癌学会総会記事 ( 日本癌学会 ) 68回 172 - 173 2009.08

Cancer epigenomics: Implications of DNA methylation in personalized cancer therapy

Minoru Toyota, Hiromu Suzuki, Toshiharu Yamashita, Koichi Hirata, Kohzoh Imai, Takashi Tokino, Yasuhisa Shinomura

CANCER SCIENCE ( WILEY ) 100 ( 5 ) 787 - 791 2009.05

Book review, literature introduction, etc.

Genetic alterations in cancer can provide information for predicting a tumor's sensitivity to chemotherapeutic drugs. But although such information is certainly useful, the relatively low frequency of mutations seen in many cancers limits the utility of pharmacogenomics in large numbers of cancer patients, necessitating consideration of other approaches. Epigenetic changes such as DNA methylation are a hallmark of human cancers. Methylation of genes involved in DNA repair and maintaining genome integrity (e.g. MGMT, hMLH1, WRN, and FANCF), and cell-cycle checkpoint genes (e.g. CHFR and 14-3-3 sigma, CDK10, and p73), all reportedly influence the sensitivity to chemotherapeutic drugs, suggesting that DNA methylation could serve as a molecular marker for predicting the responsiveness of tumors to chemotherapy. However, the comprehensive study of pharmacoepigenomics awaits the advent of genome-wide analysis of DNA methylation using microarrays and next-generation sequencers. (Cancer Sci 2009; 100: 787-791).

多発性骨髄腫におけるDNA異常メチル化の網羅的解析と,新規メチル化標的遺伝子RASD1

安井寛, 豊田実, 多羅澤功, 丸山玲緒, 野島正寛, 野島正寛, 池田博, 鈴木拓, 林敏昭, 酒井基, 石田禎夫, 麻奥英毅, 時野隆至, 今井浩三, 篠村恭久

臨床血液 49 ( 9 ) 879 2008.09

皺襞肥大型胃炎におけるゲノムワイドな低メチル化とCpGアイランド高メチル化の関連(Genome-wide hypomethylation and regional hypermethylation in H. pylori-related enlarged fold gastritis)

山本英一郎, 豊田実, 鈴木拓, 野島正寛, 丸山玲緒, 五十嵐伸一, 時野隆至, 今井浩三, 篠村恭久

日本癌学会総会記事 ( 日本癌学会 ) 67回 70 - 70 2008.09

乳癌においてDNAメチル化により不活化される遺伝子のゲノム網羅的解析(Genome-wide Analysis for the Genes Silenced by DNA Methylation in Breast Cancer)

藤兼智子, 豊田実, 鈴木拓, 西川紀子, 大村東生, 西舘敏彦, 佐々木泰史, 平田公一, 時野隆至

日本癌学会総会記事 ( 日本癌学会 ) 67回 171 - 171 2008.09

miRNA abriormalities in gastrointestinal cancers

井戸川雅史, 時野隆至, 篠村恭久

最新医学 ( 最新医学社 ) 63 ( 0 ) 1849 - 1859 2008.09

タンパクに翻訳されない21-23塩基のmRNA,microRNA(miRNA)がさまざまな遺伝子の発現を抑制していることが明らかとなってきた.がん促進的に働く発がんmiRNAやがん抑制miRNAが存在し,消化器癌でも発現の異常が認められている.一方,代表的ながん抑制遺伝子p53の標的にmiR-34ファミリーが含まれることも報告された.今後,消化器癌におけるmiRNAの役割について,さらなる成果が期待される.(著者抄録)

Frequent epigenetic inactivation of Wnt antagonist genes in breast cancer (vol 98, pg 1147, 2008)

H. Suzuki, M. Toyota, H. Carraway, E. Gabrielson, T. Ohmura, T. Fujikane, N. Nishikawa, Y. Sogabe, M. Nojima, T. Sonoda, M. Mori, K. Hirata, K. Imai, Y. Shinomura, S. B. Baylin, T. Tokino

BRITISH JOURNAL OF CANCER ( NATURE PUBLISHING GROUP ) 99 ( 2 ) 384 - 384 2008.07

Other

胃癌におけるRASSF2の異常メチル化と細胞内機能の解析

DNA異常メチル化により引き起こされる消化管の癌性糖鎖不全現象

河村由紀, 豊田実, 川島麗, 萩原輝記, 鈴木拓, 篠村恭久, 時野隆至, 今井浩三, 土肥多惠子

日本臨床分子医学会学術総会プログラム・抄録集 ( 日本臨床分子医学会 ) 45回 79 - 79 2008.07

乳癌におけるヒストンメチル化酵素PRDM 14遺伝子の診断・治療への応用の可能性

西川紀子, 豊田実, 本間敏男, 藤兼智子, 西舘俊彦, 大村東生, 今井浩三, 時野隆至, 平田公一

日本外科学会雑誌 ( (一社)日本外科学会 ) 109 ( 臨増2 ) 202 - 202 2008.04

LINE-1 Hypomethylation is Associated with Increased CpG Island Methylation in H-pylori-related Enlarged Fold Gastritis

Masanori Nojima, Eiichiro Yamamoto, Minoru Toyota, Hiromu Suzuki, Yutaka Kondo, Tamana Sanomura, Yoko Murayama, Mutsumi Ohe-Toyota, Reo Maruyama, Masami Ashida, Kyoko Fujii, Yasushi Sasaki, Norio Hayashi, Mitsuru Mori, Kohzoh Imai, Takashi Tokino, Yasuhisa Shinomura

TUMOR BIOLOGY ( KARGER ) 29 84 - 84 2008

Research paper, summary (international conference)

Epigenetic Gene Silencing and microRNA in Human Colon Cancer

Minoru Toyota, Hiromu Suzuki, Takashi Tokino, Yasuhisa Shinomura, Kohzoh Imai

TUMOR BIOLOGY ( KARGER ) 29 17 - 17 2008

Research paper, summary (international conference)

Epigenetic Silencing of Interferon Regulatory Factor(IRF) Family Genes in Colorectal and Gastric Cancer

Masaki Yamashita, Hiromu Suzuki, Masanori Nojima, Shinichi Igarashi, Yasuhisa Shinomura, Kohzoh Imai, Takashi Tokino, Fumio Itoh, Minoru Toyota

TUMOR BIOLOGY ( KARGER ) 29 43 - 43 2008

Research paper, summary (international conference)

What's New in SURGERY FRONTIER 蛋白キナーゼに関連した分子標的治療薬 EGFR阻害剤

藤兼智子, 豊田実, 時野隆至, 平田公一

Surgery Frontier ( (株)メディカルレビュー社 ) 14 ( 4 ) 430 - 432 2007.12

【遺伝子・蛋白の網羅的解析から消化器疾患に迫る】 DNAメチル化のゲノムワイドな解析

豊田実, 今井崇, 曽我部陽平, 藤兼智子, 時野隆至, 篠村恭久

分子消化器病 ( (株)先端医学社 ) 4 ( 3 ) 196 - 200 2007.09

消化器癌の発生と進展には、遺伝子の変異やゲノム構造異常だけでなく、DNAメチル化などのエピジェネティックな異常が関与する。網羅的メチル化の検出法の開発により、癌において異常メチル化している遺伝子の同定や、癌のメチル化プロファイルの解析が格段に進んだ。消化器癌において異常メチル化により不活化されている遺伝子の解析により、消化器癌の診断と治療法開発に重要な知見が得られると考えられる。(著者抄録)

Ku70とPARP-1によるbeta-catenin/TCF-4転写の制御(Regulation of the beta-catenin/TCF-4-mediated gene transactivation by Ku70 and poly(ADP-ribose) polymerase-1 (PARP-1))

井戸川雅史, 益谷美都子, 今井浩三, 広橋説雄, 篠村恭久, 時野隆至, 山田哲司

日本癌学会総会記事 ( 日本癌学会 ) 66回 235 - 235 2007.08

乳癌におけるヒストンメチル化酵素PRDM14遺伝子の発現上昇および遺伝子増幅(Gene Amplification and Overexpression of PRDM14 in Breast Cancers)

西川紀子, 豊田実, 鈴木拓, 本間敏男, 藤兼智子, 大村東生, 豊田睦美, 佐々木泰史, 園田智子, 今井浩三, 時野隆至, 平田公一

日本癌学会総会記事 ( 日本癌学会 ) 66回 477 - 477 2007.08

H.pylori感染胃炎のCDH1遺伝子メチル化解析による未分化型胃癌ハイリスク群の推定(CDH1 gene methylation profile in H. pylori-related gastritis identified high-risk group of diffuse-type gastric cancer)

山本英一郎, 豊田実, 鈴木拓, 丸山玲緒, 見田裕章, 佐々木泰史, 時野隆至, 今井浩三, 篠村恭久

日本癌学会総会記事 ( 日本癌学会 ) 66回 294 - 294 2007.08

大腸癌におけるDFNA5のエピジェネテッィクな不活性化(Epigenetic inactivation of DFNA5 in colorectal cancers)

藤兼智子, 豊田実, 鈴木拓, 丸山玲緒, 今井浩三, 平田公一, 時野隆至

日本癌学会総会記事 ( 日本癌学会 ) 66回 315 - 315 2007.08

大腸癌・胃癌におけるPRDM5のエピジェネティックな不活化(Epigenetic inactivation of PRDM5 in colorectal and gastric cancer)

山下真幸, 渡邊嘉行, 豊田実, 近藤豊, 鈴木拓, 今井崇, 丸山玲緒, 佐々木泰史, 関戸好孝, 篠村恭久, 今井浩三, 伊東文生, 時野隆至

日本癌学会総会記事 ( 日本癌学会 ) 66回 315 - 315 2007.08

Dysregulation of mitotic checkpoint proteins chfr and aurora a in HTLV-i-infected T cells

Mariko Tomita, Chie Ishikawa, Tetsuro Nakazato, Minoru Toyota, Takashi Tokino, Naoki Mori

AIDS RESEARCH AND HUMAN RETROVIRUSES ( MARY ANN LIEBERT INC ) 23 ( 4 ) 588 - 589 2007.04

Research paper, summary (international conference)

山本英一郎, 豊田実, 鈴木拓, 丸山玲緒, 見田裕章, 阿部環, 佐々木泰史, 時野隆至, 今井浩三, 篠村恭久

日本消化器病学会雑誌 ( (一財)日本消化器病学会 ) 104 ( 臨増総会 ) A160 - A160 2007.03

The positive transcription elongation factor B mediates myocyte enhancer factor 2-dependent transcription

Masanori Nojima, Hiromu Suzuki, Minoru Toyota, Takashi Tokino, Mitsuru Mori, Koh Fujinaga, Kohzoh Imai, Yasuhisa Shinomura

TUMOR BIOLOGY ( KARGER ) 28 82 - 82 2007

Research paper, summary (international conference)

癌関連機能性RNAの網羅的解析とエピジェネティックな異常

豊田実, 豊田実, 丸山玲緒, 丸山玲緒, 鈴木拓, 野島正寛, 鹿島理沙, 佐々木泰史, 今井浩三, 篠村恭久, 時野隆至

生化学 2007

p53遺伝子治療におけるヒストン脱アセチル化酵素阻害剤FK228の併用効果

佐々木泰史, 佐々木泰史, 時野隆至, 今井浩三, 篠村恭久

日本癌治療学会誌 42 ( 2 ) 2007

胃癌におけるK-ras遺伝子増幅の解析

見田裕章, 豊田実, 青木文夫, 明石浩史, 丸山玲緒, 佐々木泰史, 鈴木拓, 井戸川雅史, 鹿島理沙, SABAU Sorin, 今井浩三, 篠村恭久, 時野隆至

生化学 2007

Identification of HDGF as a transcriptional target downregulated by p53

NEGISHI Hideaki, SASAKI Yasushi, SUZUKI Hiromu, MARUYAMA Reo, TOYOTA Minoru, SAITO Tsuyoshi, TOKINO Takashi

日本癌学会学術総会記事 66th 2007

CHFR mitotic checkpoint protein inhibits the transcriptional activity of NF-kappa B

KASHIMA Lisa, MITA Hiroaki, TOYOTA Minoru, SASAKI Yasushi, SUZUKI Hiromu, IDOGAWA Masashi, TOKINO Takashi

生化学 2007

胃癌における高頻度なWntシグナル活性化とSFRP遺伝子のエピジェネティックな不活化

鈴木拓, 豊田実, 野島正寛, 丸山玲緒, 時野隆至, 篠村恭久, 森満, 今井浩三

生物物理化学 = Journal of Electrophoresis 50 ( 3 ) 128 - 128 2006.09

胃癌におけるRasシグナル関連分子のジェネティック・エピジェネティックな異常の解析

山本英一郎, 豊田実, 鈴木拓, 見田裕章, 丸山玲緒, 阿部環, 佐藤裕信, 多羅澤功, 西川紀子, 佐々木泰史, 時野隆至, 今井浩三, 篠村恭久

日本癌学会総会記事 ( 日本癌学会 ) 65回 133 - 133 2006.09

悪性胸膜中皮腫においてDNAメチル化により制御されるがん抑制遺伝子の網羅的解析

後藤康洋, 近藤豊, 横山俊彦, 谷口哲郎, 長田啓隆, 鈴木拓, 時野隆至, 豊田実, 今井浩三, 長谷川好規, 下方薫, 関戸好孝

日本癌学会総会記事 ( 日本癌学会 ) 65回 200 - 200 2006.09

大腸発癌におけるPoly(ADP-ribose)Polymerase-1(PARP-1)によるTCF/β-cateninの転写制御

井戸川雅史, 時野隆至, 篠村恭久, 今井浩三, 広橋説雄, 山田哲司

日本消化器病学会雑誌 ( (一財)日本消化器病学会 ) 103 ( 臨増大会 ) A880 - A880 2006.09

悪性リンパ腫におけるTIMP-2遺伝子のメチル化と発現抑制

多羅澤功, 鈴木拓, 豊田実, 丸山玲緒, 阿部環, 佐藤裕信, 山本英一郎, 佐々木泰史, 石田禎夫, 時野隆至, 今井浩三, 篠村恭久

日本癌学会総会記事 ( 日本癌学会 ) 65回 361 - 361 2006.09

β-cateninのFUS/TLSとの結合によるスプライシング制御

井戸川雅史, 佐藤智, 時野隆至, 篠村恭久, 今井浩三, 広橋説雄, 山田哲司

日本癌学会総会記事 ( 日本癌学会 ) 65回 38 - 39 2006.09

PARP-1とKu70の協調によるβ-catenin/TCF-4の転写制御

井戸川雅史, 時野隆至, 篠村恭久, 今井浩三, 広橋説雄, 山田哲司

日本臨床分子医学会学術総会プログラム・抄録集 ( 日本臨床分子医学会 ) 43回 61 - 61 2006.07

DNAメチル化のp53経路への関与と消化器癌における分子診断

豊田実, 丸山玲緒, 鈴木拓, 佐々木泰史, 阿部環, 佐藤裕信, 山本英一郎, 伊東文生, 時野隆至, 今井浩三, 篠村恭久

日本臨床分子医学会学術総会プログラム・抄録集 ( 日本臨床分子医学会 ) 43回 65 - 65 2006.07

胃癌におけるRasシグナル関連分子のジェネティック・エピジェネティックな異常の解析

山本英一郎, 豊田実, 鈴木拓, 見田裕章, 丸山玲緒, 阿部環, 佐藤裕信, 多羅澤功, 西川紀子, 佐々木泰史, 時野隆至, 今井浩三, 篠村恭久

日本臨床分子医学会学術総会プログラム・抄録集 ( 日本臨床分子医学会 ) 43回 71 - 71 2006.07

Comparative analysis of human and mouse genomes identified vitamin D receptor as a direct target of p53-mediated transcriptional activation

Maruyama Reo, M. Toyota, Y. Shinomura, T. Tokino, K. Imai

TUMOR BIOLOGY ( KARGER ) 27 84 - 84 2006

Research paper, summary (international conference)

The Ras effector RASSF2 is a novel tumor suppressor gene in human colorectal cancer

Minoru Toyota, A. Kimishige, T. Takashi, S. Yasuhisa, K. Imai

TUMOR BIOLOGY ( KARGER ) 27 5 - 5 2006

Research paper, summary (international conference)

消化器癌におけるリボソーム蛋白質L13の活性化と抗癌剤感受性

佐々木泰史, 小林寿久, 伊東文生, 山本博幸, 見田裕章, 豊田実, 時野隆至, 今井浩三, 篠村恭久

日本癌治療学会誌 41 ( 2 ) 2006

骨肉腫に対するp53ファミリーを用いた抗腫瘍活性の基礎的研究

大島雄一郎, 佐々木泰史, 豊田実, 見田裕章, 和田卓郎, 名越智, 川口哲, 加谷光規, 山下敏彦, 時野隆至

日本癌学会学術総会記事 65th 2006

DNAメチル化によるp53標的遺伝子のサイレンシングとがん化における意義

豊田実, 鈴木拓, 丸山玲緒, 佐々木泰史, 渡邊嘉行, 西川紀子, 阿部環, 佐藤裕信, 見田裕章, 伊東文生, 時野隆至, 今井浩三, 篠村恭久

日本癌学会学術総会記事 65th 2006

p53ファミリーを用いた骨肉腫細胞に対するアポトーシスの誘導

大島雄一郎, 大島雄一郎, 川口哲, 加谷光規, 名越智, 和田卓郎, 山下敏彦, 佐々木泰史, 豊田実, 時野隆至

日本整形外科学会雑誌 80 ( 6 ) 2006

p53ファミリー(p73,p63)の標的遺伝子flotillin-2の同定

佐々木泰史, 佐々木泰史, 大島雄一郎, 苗代康可, 苗代康可, 見田裕章, 見田裕章, 豊田実, 豊田実, 丸山玲緒, 明石浩史, 明石浩史, 篠村恭久, 今井浩三, 中村祐輔, 時野隆至

日本癌学会学術総会記事 65th 2006

p53およびvitamin D受容体(VDR)を標的とした大腸癌治療法の開発

丸山玲緒, 豊田実, 豊田実, 佐々木泰史, 明石浩史, 見田裕章, 鈴木拓, 鈴木拓, 時野隆至, 今井浩三, 篠村恭久

日本消化器病学会雑誌 103 2006

Activation of the ribosomal protein L13 gene in human gastrointestinal cancer.

KASHIMA Lisa, SASAKI Yasushi, KOBAYASHI Toshihisa, OSHIMA Yuichiro, SUZUKI Hiromu, MITA Hiroaki, TOYOTA Minoru, IMAI Kohzoh, SHINOMURA Yasuhisa, TOKINO Takashi

生化学 2006

A full genome scan for gastric cancer

The histone deacetylase inhibitor FK228 increases the efficacy of adenovirus-mediated p53 family gene therapy in in vitro and in vivo human cancer models

Y Sasaki, Y Oshima, H Mita, Y Naishiro, M Toyota, K Imai, T Tokino

JOURNAL OF CLINICAL ONCOLOGY ( AMER SOC CLINICAL ONCOLOGY ) 23 ( 16 ) 231S - 231S 2005.06

Research paper, summary (international conference)

M Aoki, Y Yamamura, H Noshiro, K Sakai, J Yokota, T Kohno, T Tokino, S Ishida, S Ohyama, Ninomiya, I, K Uesaka, M Kitajima, S Shimada, S Matsuno, M Yano, M Hiratsuka, H Sugimura, F Itoh, T Minamoto, Y Maehara, S Takenoshita, T Aikou, H Katai, K Yoshimura, T Takahashi, K Akagi, M Sairenji, K Yamamoto, T Sasazuki

JOURNAL OF MEDICAL GENETICS ( B M J PUBLISHING GROUP ) 42 ( 1 ) 83 - 87 2005.01

Detection of Chemo-responsive Fingerprints in Serum Level using Model System

土佐紀子, 土佐紀子, 苗代康可, 苗代康可, 豊田実, 豊田実, 見田裕章, 見田裕章, 佐々木泰史, 佐々木泰史, 時野隆至, 今井浩三, 小海康夫

日本プロテオーム学会大会プログラム・抄録集 2005 (Web) 2005

p53ファミリー(p53,p73,p63)の抗腫よう活性

佐々木泰史, 大島雄一郎, 見田裕章, 山下利春, 篠村恭久, 今井浩三, 時野隆至

日本癌治療学会誌 40 ( 2 ) 2005

分裂期チェックポイントCHFRの機能解析

鹿島理沙, 見田裕章, 豊田実, 豊田実, 豊田実, 佐々木泰史, 鈴木拓, 鈴木拓, 秋野公臣, 秋野公臣, 時野隆至

日本癌学会学術総会記事 64th 2005

p53ファミリーの抗腫よう活性と癌治療への応用:HDAC阻害剤FK228の併用効果も含めて

石田勢津子, 石田勢津子, 佐々木泰史, 佐々木泰史, 大島雄一郎, 見田裕章, 見田裕章, 豊田実, 豊田実, 今井浩三, 時野隆至

日本癌学会学術総会記事 64th 2005

口腔へん平上皮癌における分泌型Frizzled関連蛋白遺伝子のエピジェネティックな不活化

曽我部陽平, 曽我部陽平, 鈴木拓, 鈴木拓, 鈴木拓, 豊田実, 豊田実, 豊田実, 秋野公臣, 秋野公臣, 佐々木泰史, 見田裕章, 草野真暢, 草野真暢, 今井崇, 時野隆至, 平塚博義

日本癌学会学術総会記事 64th 2005

p53ファミリーを用いた骨肉腫に対する遺伝子治療の基礎的研究

大島雄一郎, 大島雄一郎, 佐々木泰史, 豊田実, 見田裕章, 和田貞郎, 和田貞郎, 名越哲, 名越哲, 川口哲, 川口哲, 山下敏彦, 時野隆至

日本癌学会学術総会記事 64th 2005

CpG Island Methylator Phenotype胃癌の分子生物・臨床病理学的特徴とEBウイルス関連胃癌との関係

草野真暢, 草野真暢, 豊田実, 豊田実, 鈴木拓, 鈴木拓, 秋野公臣, 秋野公臣, 見田裕章, 佐々木泰史, 垣内英樹, 伊東文生, 篠村恭久, 今井浩三, 時野隆至

日本癌学会学術総会記事 64th 2005

がん治療モデルマウスをもちいた治療応答性Chemo-responsive fingerprintsの探索

苗代康可, 土佐紀子, 土佐紀子, 豊田実, 見田裕章, 佐々木泰史, 今井浩三, 時野隆至, 小海康夫

日本癌学会学術総会記事 64th 2005

胃癌におけるメチル化標的新規癌関連遺伝子の検討

渡辺嘉行, 渡辺嘉行, 豊田実, 豊田実, 秋野公臣, 秋野公臣, 鈴木拓, 鈴木拓, 井戸川雅史, 井戸川雅史, 鹿島里沙, 見田裕章, 佐々木泰史, 伊東文生, 時野隆至

日本分子生物学会年会講演要旨集 28th 2005

p53によるビタミンD受容体の発現誘導と抗腫よう効果の増強作用

丸山玲緒, 豊田実, 豊田実, 佐々木泰史, 青木文夫, 明石浩史, 明石浩史, 見田裕章, 鈴木拓, 鈴木拓, 今井浩三, 篠村恭久, 時野隆至

日本分子生物学会年会講演要旨集 28th 2005

p53によるVDR(vitamin D receptor)の発現誘導

丸山玲緒, 豊田実, 豊田実, 明石浩史, 明石浩史, 佐々木泰史, 青木文夫, 見田裕章, 鈴木拓, 鈴木拓, 篠村恭久, 今井浩三, 時野隆至

日本癌学会学術総会記事 64th 2005

RASシグナル伝達におけるRASSF2遺伝子の機能解析

高橋文彦, 豊田実, 豊田実, 秋野公臣, 秋野公臣, 鈴木拓, 鈴木拓, 見田裕章, 佐々木泰史, 篠村恭久, 今井浩三, 時野隆至

日本癌学会学術総会記事 64th 2005

分裂期チェックポイント分子CHFRの機能解析

鹿島理沙, 見田裕章, 豊田実, 豊田実, 豊田実, 佐々木泰史, 井戸川雅史, 井戸川雅史, 鈴木拓, 秋野公臣, 秋野公臣, 時野隆至

日本分子生物学会年会講演要旨集 28th 2005

胃癌においてメチル化により不活化している新規癌抑制遺伝子の同定

上野理子, 豊田実, 豊田実, 鈴木拓, 鈴木拓, 秋野公臣, 秋野公臣, 見田裕章, 見田裕章, 佐々木泰史, 佐々木泰史, 篠村恭久, 時野隆至, 今井浩三

日本癌学会学術総会記事 64th 2005

消化器癌におけるribosomal protein L13遺伝子の活性化

佐々木泰史, 佐々木泰史, 小林寿久, 山本博幸, 伊東文生, 豊田実, 豊田実, 見田裕章, 見田裕章, 時野隆至, 今井浩三, 篠村恭久

日本分子生物学会年会講演要旨集 28th 2005

p53ファミリーp73,p63/p51の標的遺伝子の検索

佐々木泰史, 佐々木泰史, 苗代康可, 苗代康可, 見田裕章, 見田裕章, 豊田実, 豊田実, 石田勢津子, 大島雄一郎, 丸山玲緒, 明石浩史, 明石浩史, 篠村恭久, 中村祐輔, 今井浩三, 時野隆至

日本癌学会学術総会記事 64th 2005

塩基配列情報を基盤とした定量的ゲノム解析法の開発と癌研究への応用

見田裕章, 豊田実, 豊田実, 丸山玲緒, 青木文夫, 明石浩史, 佐々木泰史, 鹿島理沙, 秋野公臣, 鈴木拓, 苗代康司, 井戸川雅史, 大西浩文, 大西浩文, 辰巳治之, 今井浩三, 篠村恭久, 時野隆至

日本分子生物学会年会講演要旨集 28th 2005

胃癌において異常メチル化により不活化している新規遺伝子ACMG1

佐藤裕信, 豊田実, 豊田実, 鈴木拓, 鈴木拓, 秋野公臣, 秋野公臣, 見田裕章, 佐々木泰史, 上野理子, 篠村恭久, 篠村恭久, 時野隆至, 今井浩三

日本癌学会学術総会記事 64th 2005

アクチン結合タンパクT-fimbrinとDNA損傷に対する応答性:T-fimbrinの発現抑制は肝癌細胞株のDNA損傷に対する感受性を高める。

池田博, 佐々木泰史, 佐々木泰史, 見田裕章, 見田裕章, 豊田実, 豊田実, 篠村恭久, 時野隆至, 今井浩三

日本癌学会学術総会記事 64th 2005

5-aza-2’-deoxycytidineによるBNIP3遺伝子の発現回復はすい癌細胞における低酸素誘導性細胞死感受性を増強する

阿部環, 豊田実, 豊田実, 鈴木拓, 鈴木拓, 秋野公臣, 秋野公臣, 見田裕章, 見田裕章, 佐々木泰史, 佐々木泰史, 遠藤高夫, 篠村恭久, 時野隆至, 今井浩三

日本癌学会学術総会記事 64th 2005

ゲノム比較とニューラルネットによるp53結合配列解析システムの開発

明石浩史, 丸山玲緒, 丸山玲緒, 青木文夫, 豊田実, 豊田実, 佐々木泰史, 佐々木泰史, 見田裕章, 見田裕章, 大西浩文, 戸倉一, 苗代康司, 苗代康司, 辰巳浩之, 今井浩三, 今井浩三, 時野隆至

日本癌学会学術総会記事 64th 2005

抗がん剤応答性ペプチドフィンガープリントの網羅的検出

苗代康可, 土佐紀子, 豊田実, 見田裕章, 佐々木泰史, 今井浩三, 小海康夫, 時野隆至

日本病理学会会誌 94 ( 1 ) 2005

がん治療モデルマウスを用いた治療応答性Chemo-responsive fingerprintsの探索

土佐紀子, 苗代康可, 豊田実, 見田裕章, 佐々木泰史, 時野隆至, 今井浩三, 小海康夫

日本病理学会会誌 94 ( 1 ) 2005

Inactivation of class II transactivator by DNA methylation and histone deacetylation associated with absence of HLA-DR induction by interferon-gamma in haematopoietic tumour cells (vol 90, pg 844, 2004)

Y Morimoto, M Toyota, H Ikeda, T Tokino, K Imai

BRITISH JOURNAL OF CANCER ( NATURE PUBLISHING GROUP ) 91 ( 11 ) 1978 - 1978 2004.11

Other

統合的sequence motif解析システムの開発とその応用

明石浩史, 丸山玲緒, 青木文夫, 豊田実, 佐々木泰史, 戸倉一, 西城一翼, 山口徳蔵, 見田裕章, 苗代康可, 辰巳治之, 今井浩三, 時野隆至

医療情報学連合大会論文集 ( (一社)日本医療情報学会 ) 24回 842 - 843 2004.11

Delta Np63 isotypes differentially regulate keratinocyte proliferation and differentiation

WC Weinberg, T Yamashita, T Tokino, M Young, R Ponnamperuma, K King

JOURNAL OF INVESTIGATIVE DERMATOLOGY ( BLACKWELL PUBLISHING INC ) 122 ( 3 ) A23 - A23 2004.03

Research paper, summary (international conference)

消化管癌におけるSEZ6L遺伝子のエピジェネティックな不活化

野島正寛, 鈴木拓, 豊田実, 秋野公臣, 見田裕章, 佐々木泰史, 時野隆至, 今井浩三

日本癌学会総会記事 63rd 2004

p53ファミリー遺伝子,p63,p73によるPigment Epithelium Derived Factorの発現誘導

苗代康可, 佐々木泰史, 今井浩三, 中村祐輔, 時野隆至

日本分子生物学会年会プログラム・講演要旨集 27th 2004

ヒトとマウスのゲノム比較解析による新規p53標的遺伝子同定の試み

丸山玲緒, 豊田実, 明石浩史, 青木文夫, 佐々木泰史, 見田裕章, 垣内英樹, 辰巳治之, 時野隆至

日本癌学会総会記事 63rd 2004

消化器癌におけるMHC class II発現消失とClass II transactivatorのエピジェネティックな異常

佐藤亜由美, 豊田実, 池田英之, 鈴木拓, 見田裕章, 秋野公臣, 佐々木泰史, 時野隆至, 今井浩三

日本癌学会総会記事 63rd 2004

インシリコ解析による新規p53標的遺伝子同定の試み

丸山玲緒, 豊田実, 明石浩史, 青木文夫, 佐々木泰史, 見田裕章, 垣内英樹, 時野隆至, 今井浩三

情報計算化学生物学会大会予稿集 2004 2004

RASシグナル伝達におけるGenetic/Epigenetic異常の解析

秋野公臣, 豊田実, 鈴木拓, 草野真暢, 苗代康可, 見田裕章, 佐々木泰史, 今井浩三, 時野隆至

日本癌学会総会記事 63rd 2004

新規p53標的遺伝子としてのSCN3B遺伝子の同定

森本一郎, 足立賢哉, 豊田実, 佐々木泰史, 丸山玲緒, 日野田裕治, 工藤隆一, 今井浩三, 時野隆至

日本癌学会総会記事 63rd 2004

大腸癌における分泌型Frizzled関連蛋白遺伝子のエピジェネティックな不活化は大腸癌におけるWntシグナル経路活性化を補完する

鈴木拓, 豊田実, 野島正寛, 秋野公臣, 見田裕章, 佐々木泰史, 苗代康可, 時野隆至, 森満

日本癌学会総会記事 63rd 2004

p53結合配列解析のための統合的解析システムの開発

明石浩史, 丸山玲緒, 青木文夫, 豊田実, 佐々木泰史, 見田裕章, 苗代康可, 辰巳治之, 時野隆至

日本癌学会総会記事 63rd 2004

抗がん剤応答性ペプチドフィンガープリントの網羅的検出

苗代康可, 土佐紀子, 豊田実, 見田裕章, 佐々木泰史, 小海康夫, 今井浩三, 時野隆至

日本癌学会総会記事 63rd 2004

ヒトとマウスのゲノム比較解析による新規p53標的遺伝子の同定

丸山玲緒, 豊田実, 明石浩史, 青木文夫, 佐々木泰史, 見田裕章, 垣内英樹, 辰巳治之, 時野隆至

日本分子生物学会年会プログラム・講演要旨集 27th 2004

p73転写制御因子によるIL-4シグナルの活性化

佐々木泰史, 見田裕章, 豊田実, 石田勢津子, 山下利春, 田中敏博, 中村祐輔, 今井浩三, 時野隆至

日本癌学会総会記事 63rd 2004

造血器腫ようにおけるアポトーシス関連遺伝子のエピジェネティックな異常の解析

高橋文彦, 豊田実, 村井政史, 鈴木拓, 見田裕章, 佐々木泰史, 石田禎夫, 時野隆至, 今井浩三

日本癌学会総会記事 63rd 2004

アクチン結合蛋白T-fimbrinによるG2/M期制御と抗癌剤感受性

佐々木泰史, 池田博, 見田裕章, 豊田実, 時野隆至, 日野田裕治, 今井浩三

日本分子生物学会年会プログラム・講演要旨集 27th 2004

ヒト胃癌におけるナトリウムトランスポーターファミリー遺伝子SLC5A8の異常メチル化と発現抑制

上野理子, 豊田実, 秋野公臣, 見田裕章, 村井政史, 佐々木泰史, 鈴木拓, 遠藤高夫, 時野隆至

日本癌学会総会記事 63rd 2004

Berrant methylation and gene silencing of proapoptotic genes, hrk, in gastrointestinal tumors.

F Itoh, M Toyota, T Obata, A Satoh, Y Sasaki, H Suzuki, T Tokino, JPJ Issa, K Imai

CLINICAL CANCER RESEARCH ( AMER ASSOC CANCER RESEARCH ) 9 ( 16 ) 6160S - 6160S 2003.12

Research paper, summary (international conference)

End User指向ネットワーク対応型ゲノム解析システムの開発とその応用によるp53 family gene標的遺伝子の同定

明石浩史, 豊田実, 青木文夫, 佐々木泰史, 大西浩文, 見田裕章, 戸倉一, 苗代康可, 丸山玲緒, 中村正弘, 今井浩三, 時野隆至, 辰巳治之, 秋野豊明

医療情報学連合大会論文集 ( (一社)日本医療情報学会 ) 23回 439 - 440 2003.11

Histone deacetylation, but not hypermethylation, modifies class II transactivator and MHC class II gene expression in squamous cell carcinomas.

Takayuki Kanaseki, Hideyuki Ikeda, Yukio Takamura, Minoru Toyota, Yoshihiko Hirohashi, Takashi Tokino, Tetsuo Himi, Noriyuki Sato

Journal of immunology (Baltimore, Md. : 1950) 170 ( 10 ) 4980 - 5 2003.05 [Refereed] [International journal]

In this study, we first categorized nine squamous cell carcinoma (SCC) cell lines into two groups in terms of the expression of HLA-DR, -DP, and -DQ molecules. Subsequently, the expression of class II transactivator (CIITA) was studied in these cell lines, because it is widely accepted that the expression of MHC class II molecules is regulated by different types of CIITA transcripts that are initiated by distinct promoters. The majority of the SCC cell lines (six of nine) expressed HLA-DR molecules and CIITA promoter IV (pIV) transcripts in the presence of IFN-gamma. In contrast, three of the nine SCC cell lines were completely negative for class II molecules and all types of CIITA, suggesting epigenetic changes in the promoter region in these cells. Previously, methylation of CIITA pIV was reported to silence CIITA gene expression. We extensively studied the methylation status of CIITA pIV using a panel of 22 SCC cell lines. Remarkably, none of the SCC cell lines demonstrated hypermethylation at the site. In contrast, treatment with a histone deacetylation inhibitor in combination with IFN-gamma clearly restored the expression of the CIITA type IV gene in the HLA-DR-negative SCC cell lines, and the acetylation status of histone H3 examined by chromatin immunoprecipitation analysis was closely associated with the gene expression. Moreover, stable transfection of the CIITA gene into an HLA-DR-negative cell line restored constitutive expression of MHC class II molecules. Therefore, histone deacetylation, but not hypermethylation, modifies CIITA DNA and class II gene expression in SCC.

Bioinformaticsによるゲノム・遺伝子解析と臨床への応用

明石浩史, 豊田実, 時野隆至, 辰巳治之, 今井浩三

Drug delivery system 18 ( 3 ) 218 - 218 2003.05

Analysis of lung tumorigenesis in chimeric mice indicates the Pulmonary adenoma resistance 2 (Par2) locus to operate in the tumor-initiation stage in a cell-autonomous manner: detection of polymorphisms in the Poli gene as a candidate for Par2.

Gang-Hong Lee, Hiroyuki Nishimori, Yasushi Sasaki, Hiroshi Matsushita, Tomoyuki Kitagawa, Takashi Tokino

Oncogene 22 ( 15 ) 2374 - 82 2003.04 [Refereed] [International journal]

The Pulmonary adenoma resistance 2 (Par2) locus of the BALB/cByJ mouse, located within 0.5 cM of chromosome 18, is responsible for reducing the mean multiplicity of urethane-induced lung tumors relative to those in C57BL/6J, A/J and C3H/HeJ mice. Thus, BALB/B6-Par2 congenic strain genetically identical to BALB/cByJ except carrying C57BL/6J Par2 alleles develops seven times more tumors than BALB/cByJ. To gain clues for identification of Par2 candidate genes, we analysed lung tumorigenesis in BALB/cByJ<-->BALB.B6-Par2 chimeric animals. Of 100 tumors induced by urethane in 16 chimeras, 82 originated from BALB.B6-Par2 cells, indicating the Par2 phenotype to be cell-autonomous. In addition, the BALB.B6-Par2- and BALB/cByJ-derived tumors were similar in mean size, implying that the phenotype is primarily expressed during initiation rather than in the promotion stage of carcinogenesis. Given these results, we surveyed a comprehensive mouse genome database and physically mapped Par2 within a 2.3 Mbp segment containing three known genes, Poli, Mbd2 and Dcc. Among those, the Poli seemed to be the most reasonable Par2 candidate, since it encodes an extremely error-prone DNA polymerase preferentially incorporating G or T opposite template T in vitro, reminiscent of the Kras2 activation because of an A to G or T point mutation within codon 61 with which most urethane-induced lung tumors are initiated. Indeed, our sequencing of Poli cDNAs from BALB/cByJ, C57BL/6J, A/J and C3H/HeJ lungs revealed 21 BALB/cByJ-specific single-nucleotide polymorphisms in the coding region accompanied by seven amino-acid substitutions and an elevated frequency of alternative splicing, while no polymorphisms associated with tumor susceptibility were found for either Mbd2 or Dcc. Notably, we obtained evidence that BALB/cByJ Par2 alleles may selectively decrease the frequency of Kras2-mutated tumors compared with C57BL/6J alleles. Consequently, the Poli is an intriguing Par2 candidate clearly deserving further evaluation.

Induction of tenascin-C by tumor-specific EWS-ETS fusion genes.

Goichi Watanabe, Hiroyuki Nishimori, Hideto Irifune, Yasushi Sasaki, Setsuko Ishida, Hitoshi Zembutsu, Toshihiro Tanaka, Satoshi Kawaguchi, Takuro Wada, Jun-ichi Hata, Moriaki Kusakabe, Koichi Yoshida, Yusuke Nakamura, Takashi Tokino

Genes, chromosomes & cancer 36 ( 3 ) 224 - 32 2003.03 [Refereed] [International journal]

Ewing sarcoma (ES) and peripheral primitive neuroectodermal tumors (PNETs) are associated with a chromosomal translocation resulting in a fusion of the amino-terminus of EWS with the DNA-binding domain of an ETS transcription factor (most commonly FLI1 or ERG). Although previous reports suggested that these chimera proteins would act as aberrant transcription factors, their downstream targets have not been fully elucidated. To identify downstream targets of these EWS-ETS fusion proteins, we introduced EWS-ETS fusion constructs into a human fibrosarcoma cell line, HT-1080, by retroviral transduction. Here we report that Tenascin-C (TNC) is induced to a significantly higher level in cells expressing EWS-ETSs than in cells expressing normal ETSs. Furthermore, through use of an antisense cDNA expression vector we show that expression of endogenous TNC mRNA and protein were reduced coordinately with attenuation of EWS-FLI1 fusion protein expression. A chromatin immunoprecipitation assay showed direct interaction between the TNC promoter and the EWS-FLI1 fusion protein in vivo. In addition, a luciferase reporter assay revealed that EWS-ETSs upregulated the TNC gene through four ETS binding sites in the TNC promoter. High levels of TNC expression were observed in a subset of ES cell lines (3 of 6) and primary tumors (4 of 6). Together with previous studies showing that TNC expression is involved in the invasive and malignant phenotype of several tumor types, our data suggest that the oncogenic effect of EWS-ETS may be mediated in part by upregulating of TNC expression.

DNA損傷により発現誘導されるタンパクt-fimbrinのG2/Mcheckpointへの関与と抗癌剤感受性

池田博, 佐々木泰史, 豊田実, 見田裕章, 時野隆至, 今井浩三

日本癌治療学会誌 38 ( 2 ) 2003

ヒト胃癌におけるナトリウムトランスポーターファミリー遺伝子SLC5A8の異常メチル化と発現抑制

田賀理子, 豊田実, 秋野公臣, 佐藤亜由美, 見田裕章, 村井政史, 佐々木泰史, 鈴木拓, 時野隆至

日本分子生物学会年会プログラム・講演要旨集 26th 2003

p53ファミリーの抗腫よう活性と癌治療への応用

佐々木泰史, 石田勢津子, 苗代康可, 今井浩三, 時野隆至

日本癌学会総会記事 62nd 2003

アクチン結合タンパクT-fimbrinの遺伝子発現と抗癌剤感受性との関連性

池田博, 佐々木泰史, 小林寿久, 長谷川公子, 見田裕章, 豊田実, 時野隆至, 今井浩三

日本消化器病学会雑誌 100 2003

新しいユビキチンリガーゼCHFRの不活化とM期チェックポイント制御における役割

佐藤亜由美, 豊田実, 佐々木泰史, 見田裕章, 荻和弘, 鈴木拓, 遠藤高夫, 伊東文生, 時野隆至

日本癌学会総会記事 62nd 2003

ヒト腫ようにおけるM期チェックポイント遺伝子CHFRの発現抑制と微小管阻害剤感受性

佐藤亜由美, 豊田実, 見田裕章, 佐々木泰史, 秋野公臣, 田賀理子, 丸山玲緒, 遠藤高夫, 時野隆至

日本分子生物学会年会プログラム・講演要旨集 26th 2003

p53ファミリー遺伝子の抗腫よう活性

時野隆至, 佐々木泰史

日本癌治療学会誌 38 ( 2 ) 2003

口腔へん平上皮癌におけるM期チェックポイント遺伝子の発現異常とmicrotubule inhibitor感受性

荻和弘, 豊田実, 佐々木泰史, 見田裕章, 佐藤亜由美, 今井浩三, 時野隆至

日本癌治療学会誌 38 ( 2 ) 2003

インシリコゲノム解析を利用したp53標的遺伝子の同定と応用

丸山玲緒, 豊田実, 佐々木泰史, 明石浩史, 見田裕章, 秋野公臣, 辰巳治之, 時野隆至, 今井浩三

日本分子生物学会年会プログラム・講演要旨集 26th 2003

p53ファミリーの抗腫よう活性と癌治療への応用

佐々木泰史, 苗代康可, 今井浩三, 時野隆至

日本消化器病学会雑誌 100 2003

消化器癌におけるTCF2遺伝子のエピジェネティックな異常

丸山玲緒, 豊田実, 荻和弘, 鈴木拓, 秋野公臣, 佐々木泰史, 見田裕章, 佐藤亜由美, 時野隆至

日本癌学会総会記事 62nd 2003

p53ファミリーの抗腫よう活性と癌治療への応用

佐々木泰史, 苗代康可, 今井浩三, 山下利春, 時野隆至

日本癌治療学会誌 38 ( 2 ) 2003

全ゲノム配列を対象とするsequence motif解析システムの開発と臨床応用

明石浩史, 豊田実, 青木文夫, 佐々木泰史, 見田裕章, 辰巳治之, 時野隆至, 今井浩三

日本癌学会総会記事 62nd 2003

M期チェックポイント遺伝子CHFRの機能解析

見田裕章, 豊田実, 佐藤亜由美, 荻和弘, 佐々木泰史, 苗代康可, 秋野公臣, 今井浩三, 時野隆至

日本分子生物学会年会プログラム・講演要旨集 26th 2003

p53ファミリーのアポトーシス誘導活性と癌治療への応用

佐々木泰史, 時野隆至, 今井浩三

日本内科学会雑誌 92 2003

【p53経路とRB経路ここまでわかった細胞癌化のメカニズム】 p53ファミリーの新しい機能

佐々木泰史, 時野隆至

細胞工学 ( (株)学研メディカル秀潤社 ) 22 ( 1 ) 34 - 38 2002.12

p53ファミリー遺伝子はp53と高い相同性を有するが,ヒト癌における遺伝子変異の頻度は低く,p53ファミリーメンバーの生体での役割はp53自身とは異なることが予想される.cDNAマイクロアレイ解析によって,Notch受容体のリガンドをコードするJAG1及びJAG2遺伝子がp63及びp73によって特異的に発現上昇することを見つけた.p63遺伝子の機能の少なくとも一部はJAG1/2を介して細胞,組織の分化・発生に深く関与していることが明らかになってきた.p53依存性アポトーシスにおけるp53ファミリーメンバーの役割について言及した

The Id2 gene is a novel target of transcriptional activation by EWS-ETS fusion proteins in Ewing family tumors.

Hiroyuki Nishimori, Yasushi Sasaki, Koichi Yoshida, Hideto Irifune, Hitoshi Zembutsu, Toshihiro Tanaka, Tomoki Aoyama, Taisuke Hosaka, Satoshi Kawaguchi, Takuro Wada, Jun-Ichi Hata, Junya Toguchida, Yusuke Nakamura, Takashi Tokino

Oncogene 21 ( 54 ) 8302 - 9 2002.11 [Refereed] [International journal]

We report here that the Id2 (inhibitor of DNA binding 2) gene is a novel target of transcriptional activation by EWS-FLI1 and EWS-ERG, two fusion proteins that characterize Ewing family tumors (EFTs). To identify downstream targets of these EWS-ETS fusion proteins, we introduced EWS-ETS fusion constructs into a human fibrosarcoma cell line by retroviral transduction. cDNA microarray analysis revealed that Id2 expression was up-regulated by introducing the EWS-ETS fusion gene but not by the normal full-length ETS gene. An Id2 promoter-luciferase reporter assay showed that transactivation by EWS-ETS involves the minimal Id2 promoter and may function in cooperation with c-Myc within the full-length regulatory region. A chromatin immunoprecipitation assay revealed direct interaction between the Id2 promoter and EWS-FLI1 fusion protein in vivo. Significantly higher expression of Id2 and c-Myc was observed in all of the six EFT cell lines examined compared to six other sarcoma cell lines. Moreover, high levels of Id2 expression were also observed in five of the six primary tumors examined. Id2 is generally thought to affect the balance between cell differentiation and proliferation in development and is highly expressed in several cancer types. Considering these previous studies, our data suggest that the oncogenic effect of EWS-ETS may be mediated in part by up-regulating Id2 expression. doi:10.1038/sj.onc.1206025

高速並列処理技術を用いたウェブベース全ゲノム解析システムの開発とその評価

明石浩史, 青木文夫, 豊田実, 佐々木泰史, 中村正弘, 大西浩文, 戸倉一, 西城一翼, 伊東文生, 今井浩三, 時野隆至, 辰巳治之

医療情報学連合大会論文集 ( (一社)日本医療情報学会 ) 22回 252 - 253 2002.11

DNA methylation changes in gastrointestinal disease.

Minoru Toyota, Fumio Itoh, Takefumi Kikuchi, Ayumi Satoh, Toshiro Obata, Hiromu Suzuki, Suguru Ishii, Takao Endo, Takashi Tokino, Kohzoh Imai

Journal of gastroenterology 37 Suppl 14 97 - 101 2002.11 [Refereed] [Domestic journal]

DNA methylation of the 5' region of genes is often associated with gene silencing in X-chromosome inactivation and imprinting. Recent studies have indicated that altered DNA methylation plays a role in the inactivation of multiple tumor suppressor genes and DNA repair genes such as p16INK4A and hMLH1. Colorectal adenomas have a relatively high frequency of methylation, and aberrant methylation is an early event during tumorigenesis. In aging patients, even colon epithelium which appears to be normal showed a significant amount of methylation in a subset of the genes. Colon mucosa from patients with inflammatory bowel disease also showed a high level of methylation. DNA methylation can be a specific diagnostic marker in gastrointestinal cancer and inflammatory bowel disease, for which there is no perfect marker for a noninvasive diagnosis.

Aberrant methylation of multiple genes and clinicopathological features in oral squamous cell carcinoma.

Kazuhiro Ogi, Minoru Toyota, Mutsumi Ohe-Toyota, Nobuyuki Tanaka, Makoto Noguchi, Tomoko Sonoda, Geniku Kohama, Takashi Tokino

Clinical cancer research : an official journal of the American Association for Cancer Research 8 ( 10 ) 3164 - 71 2002.10 [Refereed] [International journal]

The purpose of this study was to examine the methylation profile of various oral squamous cell carcinomas and to correlate the methylation of particular chromosomal loci with the clinicopathological features of the tumors. A semiquantitative analysis of the methylation status of 12 loci in 96 primary tumors and 13 cell lines was carried out. Methylation frequency was calculated as the percentage of methylated alleles detected by bisulfate-PCR. Of the 12 loci examined, 9 (p16INK4A, p15INK4B, p14ARF, DCC, DAP kinase, MINT1, MINT2, MINT27, and MINT31) exhibited aberrant methylation at various frequencies, whereas 3 (hMLH1, HRK, and CACNA1G) showed no methylation. Dense methylation of the 5' CpG island of DAP kinase and MINT1 was well correlated with loss of gene expression. In addition, methylation of DCC was correlated with bone invasion by gingival tumors (P = 0.036), with aggressive invasiveness of tumors of the tongue (P = 0.046), and with reduced survival (P = 0.050). Methylation of MINT1 and MINT31 also correlated with poor prognoses (P = 0.058 and 0.041), whereas methylation of p14ARF correlated with a good prognosis (P = 0.021). Cox regression analysis showed methylation of MINT31 to be an independent predictor of outcome (hazard ratio, 3.79; 95% confidence interval, 1.58-9.10) and to be associated with the T4 disease group (hazard ratio, 5.71; 95% confidence interval, 1.25-26.07). Analysis of DNA methylation is a useful approach to evaluation of the biological characteristics of oral cancers and may be a useful diagnostic indicator of patient prognosis.

DNA methylation and histone deacetylation associated with silencing DAP kinase gene expression in colorectal and gastric cancers

A Satoh, M Toyota, F Itoh, T Kikuchi, T Obata, Y Sasaki, H Suzuki, A Yawata, M Kusano, M Fujita, M Hosokawa, K Yanagihara, T Tokino, K Imai

BRITISH JOURNAL OF CANCER ( NATURE PUBLISHING GROUP ) 86 ( 11 ) 1817 - 1823 2002.06

Death-associated protein kinase is a positive regulator of programmed cell death induced by interferon, To investigate the role of epigenetic inactivation of death-associated protein kinase in gastrointestinal cancer, we examined the methylation status of the 5' CpG island of the death-associated protein kinase gene. Methylation of the 5' CpG island was detected in 3 of 9 colorectal and 3 of 17 gastric cancer cell lines, while among primary tumours, it was detected in 4 of 28 (14%) colorectal and 4 of 27 (15%) gastric cancers. By contrast, methylation of the edge of the CpG island was detected in virtually every sample examined. Death-associated protein kinase expression was diminished in four cell lines that showed dense methylation of the 5' CpG island, and treatment with 5-aza-2'-deoxycitidine, a methyltransferase inhibitor, restored gene expression, Acetylation of histones H3 and H4 in the 5' region of the gene was assessed by chromatin immunoprecipitation and was found to correlate directly with gene expression and inversely with DNA methylation, Thus, aberrant DNA methylation and histone deacetylation of the 5' CpG island, but not the edge of the CpG island, appears to play a key role in silencing death-associated protein kinase expression in gastrointestinal malignancies.

Inactivation of p57KIP2 by regional promoter hypermethylation and histone deacetylation in human tumors.

Takefumi Kikuchi, Minoru Toyota, Fumio Itoh, Hiromu Suzuki, Toshiro Obata, Hiroyuki Yamamoto, Hideki Kakiuchi, Masanobu Kusano, Jean-Pierre J Issa, Takashi Tokino, Kohzoh Imai

Oncogene 21 ( 17 ) 2741 - 9 2002.04 [Refereed] [International journal]

To clarify the role of DNA methylation in the silencing of the expression of cyclin-dependent kinase inhibitor p57KIP2 seen in certain tumors, we investigated the methylation status of its 5' CpG island in various tumor cell lines and primary cancers. Dense methylation of the region around the transcription start site was detected in 1 out of 10 colorectal, 2 out of 8 gastric, and 6 out of 14 hematopoietic tumor cell lines and in 5 out of 35 (14%) gastric, 6 out of 20 (30%) hepatocellular, and 2 out of 18 (11%) pancreatic cancers; 7 out of 25 (28%) acute myeloid leukemia cases also showed methylation of the p57KIP2 gene, which strongly correlated with the CpG island methylator phenotype (P<0.001). Detailed mapping revealed that dense methylation of the region around the transcription start site (-300 to +400), but not of the edges of the CpG island, was closely associated with gene silencing. 5-aza-2'-deoxycytidine, a methyltransferase inhibitor, restored expression of p57KIP2, and chromatin immunoprecipitation using anti-histone H3 and H4 antibodies showed histone to be deacetylated in cell lines where p57KIP2 was methylated at the transcription start site. Regional methylation and histone deacetylation thus appear to be crucially involved in the silencing of p57KIP2 expression in human tumors.

Detection of human papillomavirus DNA by PCR/microfluorometry for screening of cervical cancer.

Akihiro Yamaguchi, Naoko Hashimoto, Wataru Tsutae, Kuniyoshi Seino, Yasuhiko Ebina, Takashi Tokino, Noriyuki Sato, Kokichi Kikuchi

Clinica chimica acta; international journal of clinical chemistry 318 ( 1-2 ) 41 - 9 2002.04 [Refereed] [International journal]

BACKGROUND: Cervical cancer screening is conducted by a cytological Papanicolaou (Pap) test. For screening, it is becoming increasingly important to introduce a more objective result, based on human papillomavirus (HPV) DNA test. We describe here a practical method allowing the mass detection of HPV-DNA by PCR followed by fluorogenic DNA intercalation. METHODS: Samples used were cervical scrapes or biopsy specimens obtained from women who had undergone cytological testing for cervical cancer. Crude DNAs were extracted by a simplified proteinase K-boil method. Common and type-specific primers were newly designed for major types of high-risk HPVs. A fluorogenic DNA intercalator, SYBR Green I was directly added to the specific PCR products. The resultant fluorescence was measured by a conventional fluorometric microplate reader. RESULTS: The proposed PCR/microfluorometry (MFL) allowed a simple, rapid and economical detection of HPV-DNA without any use of labeling primers or probes. HPV-DNAs were found in 48.2% (123/255) of the cervical scrapes. The detection rate of HPV in cervical cancer biopsy specimen was 92.4% (61/66). CONCLUSIONS: PCR/MFL detection of HPV-DNA, followed by combined type-specific PCR, is expected to be an extremely useful tool in cervical cancer screening.

p53ファミリー遺伝子,p63とp73の個性

佐々木泰史, 時野隆至

最新医学 ( (株)最新医学社 ) 57 ( 4 ) 947 - 951 2002.04

Identification of the osteopontin gene as a direct target of TP53.

Ichiro Morimoto, Yasushi Sasaki, Setsuko Ishida, Kohzoh Imai, Takashi Tokino

Genes, chromosomes & cancer 33 ( 3 ) 270 - 8 2002.03 [Refereed] [International journal]

The TP53 tumor suppressor gene regulates a number of genes that are involved in cell-cycle inhibition, apoptosis, and maintaining genetic stability. Recently, two genes that have a role in immunosurveillance were identified as downstream targets of TP53. These genes, TAP1 and fractalkine, may contribute to suppress tumor growth through host immunosurveillance. It has been reported that the mouse secreted phosphoprotein osteopontin (Opn) is one of the key cytokines for type 1 immune responses mediated by macrophages. It also was reported that Opn may play a role in suppressing tumor growth in vivo. Here we identified Opn as a Tp53-target gene using mRNA differential display analysis of embryonic fibroblasts from Tp53-deficient mice. Furthermore, we found that Opn expression was upregulated by DNA damage-induced Tp53 activity and by adenovirus-mediated transfer of the human TP53 gene. In addition, a luciferase assay showed that the Opn gene has a functional Tp53-responsive element in its promoter region, and a chromatin immunoprecipitation assay confirmed interaction between the Opn promoter and Tp53 protein in vivo. These results suggest that OPN is a direct transcriptional target of TP53. The TP53-directed regulation of OPN expression suggests a novel model of TP53 participation in immunosurveillance, involving interaction with the host immune system to prevent damaged cells from undergoing malignant transformation.

The p53 family member genes are involved in the Notch signal pathway.

Yasushi Sasaki, Setsuko Ishida, Ichiro Morimoto, Toshiharu Yamashita, Takashi Kojima, Chikashi Kihara, Toshihiro Tanaka, Kohzoh Imai, Yusuke Nakamura, Takashi Tokino

The Journal of biological chemistry 277 ( 1 ) 719 - 24 2002.01 [Refereed] [International journal]

The p53 tumor suppressor is a transcription factor that regulates cell growth and death in response to environmental stimuli such as DNA damage. p63/p51 and p73 were recently identified as members of the p53 gene family. In contrast to p53 however, p63 and p73 are rarely mutated in human cancers. Mice that lack p53 are developmentally normal, while p63 and p73 appear to play critical roles in normal development. To determine how p63 and p73 are involved in normal development, we attempted to identify target genes that are specifically regulated by p63 and/or p73 but not by p53. We found that the Jagged1 (JAG1) and Jagged2 (JAG2) genes, encoding ligands for the Notch receptors, are up-regulated by p63 and p73. Furthermore, we identified a p63-binding site in the second intron of the JAG1 gene, which can directly interact with the p63 protein in vivo, as assessed by a chromatin immunoprecipitation assay. A heterologous reporter assay revealed that this p63-binding site is a functional response element and is specific for p63. We also found a target of Notch signaling, HES-1 was up-regulated in Jurkat cells, in which Notch1 is highly expressed, when co-cultured with p63-transfected cells, suggesting that p63 can trigger the Notch signal pathway in neighboring cells. Our findings show an association between the p53 family genes and Notch signaling and suggest a potential molecular mechanism for the involvement of the p53 family genes in normal development.

新しいp53標的遺伝子SCBP1の同定と機能解析

足立賢哉, 豊田実, 佐々木泰史, 石田勢津子, 斉藤豪, 工藤隆一, 時野隆至

日本癌学会総会記事 61st 2002

DNAメチル化による遺伝子不活化の分子機構

豊田実, 菊地剛史, 佐藤亜由美, 佐々木泰史, 小畑俊郎, 秋野公臣, 鈴木拓, 時野隆至, 今井浩三

日本癌学会総会記事 61st 2002

消化器癌におけるアポトーシス関連遺伝子Hrkの異常メチル化

小畑俊郎, 豊田実, 佐々木泰史, 佐藤亜由美, 菊地剛史, 鈴木拓, 伊東文生, 時野隆至, 今井浩三

日本癌学会総会記事 61st 2002

p53ファミリー遺伝子,p63/p51,p73によるNotchシグナルの活性化

佐々木泰史, 石田勢津子, 森本一郎, 田中敏博, 今井浩三, 中村祐輔, 時野隆至

日本癌学会総会記事 61st 2002

p53ファミリーを用いた消化器癌に対する遺伝子治療の基礎的研究 HDAC阻害剤FK228の併用効果も含めて

佐々木泰史, 今井浩三, 時野隆至

日本癌治療学会誌 37 ( 2 ) 2002

全ゲノムを対象とする高速並列処理によるp53結合部位解析システムの開発とその臨床応用

明石浩史, 豊田実, 青木文夫, 佐々木泰史, 西森博幸, 今井浩三, 辰巳治之, 時野隆至

日本癌学会総会記事 61st 2002

【ゲノム医療と情報システム】情報システムのポストゲノムプロジェクトへの応用

辰巳治之, 青木文夫, 明石浩史, 宮司正道, 豊田実, 佐々木泰史, 西森博幸, 時野隆至, 伊東文生, 今井浩三

新医療 ( (株)エムイー振興協会 ) 28 ( 11 ) 64 - 69 2001.11

ネットワーク・マルチパラレル・コンピューティングによる遺伝子解析システムの開発

青木文夫, 明石浩史, 豊田実, 宮司正道, 佐々木泰史, 西森博幸, 時野隆至, 辰巳治之

医療情報学連合大会論文集 ( (一社)日本医療情報学会 ) 21回 534 - 535 2001.11

Induction of apoptosis in melanoma cell lines by p53 and its related proteins

T Yamashita, T Tokino, H Tonoki, T Moriuchi, HY Jin, F Omori, K Jimbow

JOURNAL OF INVESTIGATIVE DERMATOLOGY ( BLACKWELL SCIENCE INC ) 117 ( 4 ) 914 - 919 2001.10

Melanoma cells rarely contain mutant p53 and hardly undergo apoptosis by wild-type p53. By using recombinant adenoviruses that express p53 or p53-related p51A or p73 beta, we tested their apoptotic activities in melanoma cells. Yeast functional assay revealed a mutation of p53 at the 258th codon (AAA [K] instead of GAA [E]) in one cell line, 70W, out of six human melanoma cell fines analyzed (SK-mel-23, SK-mel-24, SK-mel-118, TXM18, 70W, and G361). Adenovirus-mediated transfer of p53, p51A, and/or p73 beta suppressed growth and induced apoptotic DNA fragmentation of SK-mel-23, SK-mel-118, and 70W cells. Interestingly, p51A induced DNA fragmentation in them more significantly than p53 and p73 beta. Western blotting we analyzed levels of apoptosis-related proteins in cells expressing p53 family members. Apoptotic Bax and antiapoptotic Bcl-2 were not significantly upregulated or downregulated by expression of p53, p51A, or p73 beta, except for p53-expressing 70W cells, which contained a larger amount of Bax protein than LacZ-expressing cells. Activation of caspase-3 was demonstrated only in p51A-expressing SK-mel-118 cells. We show here that p51A can mediate apoptosis in both wild-type and mutant p53-expressing melanoma cells more significantly than p53 and p73 beta. It is also suggested that in melanoma cells (i) cellular target protein(s) other than Bcl-2 and Bax might be responsible for induction of p51A-mediated apoptosis and (ii) caspase-3 is not always involved in the apoptosis by p53 family members.

Adenovirus-mediated transfer of the p53 family genes, p73 and p51/p63 induces cell cycle arrest and apoptosis in colorectal cancer cell lines: potential application to gene therapy of colorectal cancer

Y Sasaki, Morimoto, I, S Ishida, T Yamashita, K Imai, T Tokino

GENE THERAPY ( NATURE PUBLISHING GROUP ) 8 ( 18 ) 1401 - 1408 2001.09

p53 gene therapy is being tested clinically for the treatment of human cancer, however, some cancer models (in vivo and in vitro) are resistant to p53. To explore the potential use of two p53 homologues, p73 and p51/p63, in cancer gene therapy, we introduced p53, p73 and p51/p63 into colorectal cancer cell lines via adenoviral vectors, and compared their effects on cell growth. Among 10 cell lines tested, six cell lines displayed a similar response following transduction of p53, p73 beta or p51A/p63 gamma; two lines underwent cell-cycle arrest, three lines exhibited apoptosis and one line showed no-effect following transduction. The effect on cell-cycle progression was variable in the other four cell lines. Interestingly, three cell lines were resistant to p53-mediated apoptosis, including two lines having endogenous wild-type p53 alleles, but underwent apoptosis after transduction of p73 beta or p51A/p63 gamma. Similar to p53, transduction of p51A/p63 gamma induced extensive apoptosis when combined with adriamycin or X-radiation in SW480 cells, which are normally resistant to apoptosis. Transduction of p73 beta and p51A/p63 gamma also reduced the tumorigenicity of two colorectal cancer cells in vivo. These results suggest that adenovirus-mediated p73 beta and p51A/p63 gamma transfer are potential novel approaches for the treatment of human cancers, particularly for tumors that are resistant to p53 gene therapy.

アデノウイルスベクターを用いたp53ファミリー遺伝子導入は大腸癌由来細胞株に細胞周期停止とアポトーシスを誘導する大腸癌の遺伝子治療への応用

佐々木泰史, 時野隆至

放射線生物研究 ( 放射線生物研究会 ) 36 ( 3 ) 279 - 290 2001.09

【ネットワーク最前線】ポストゲノム・プロジェクトのアプリケーション p53遺伝子機能解析とマルチパラレルコンピューティング

青木文夫, 辰巳治之, 明石浩史, 宮司正道, 豊田実, 佐々木泰史, 西森博幸, 時野隆至

医療とコンピュータ ( (株)日本電子出版 ) 12 ( 9 ) 31 - 40 2001.09

Contribution of caspase-3 differs by p53 status in apoptosis induced by X-irradiation

D Kobayashi, T Tokino, N Watanabe

JAPANESE JOURNAL OF CANCER RESEARCH ( BUSINESS CENTER ACADEMIC SOCIETIES JAPAN ) 92 ( 4 ) 475 - 481 2001.04

We investigated the effect of p53 status on involvement of caspase-3 activation in cell death induced by X-irradiation, using rat embryonic fibroblasts (REFs) transduced with a temperature-sensitive mutant (mt) p53 gene. Cells with wild-type (wt) p53 showed greater resistance to X-irradiation than cells with mt p53. In cells with wt p53, X-irradiation-induced apoptosis was not inhibited by the caspase-3 inhibitor acetyl-L-aspartyl-L-methionyl-L-glutaminyl-L-aspartyl-aldehyde (Ac-DMQD-CHO) and caspase-3 activity was not elevated following X-irradiation, although induction of p53 and p21/WAF-1 protein was observed. In contrast, irradiated cells with mt p53 showed 89% inhibition of cell death with Ac-DMQD-CHO and 98% inhibition with the antioxidant N-acetyl-L-cysteine (NAC), In cells with mt p53, caspase-3 activity was increased approximately 5 times beyond baseline activity at 24 h after irradiation. This increase was almost completely inhibited by NAG. However, inhibition of caspase-3 by Ac-DMQD-CHO failed to decrease production of reactive oxygen species by cells with mt p53. Differential involvement of caspase-3 is a reason for differences in sensitivity to X-irradiation in cells with different p53 status. Caspase-3 activation appears to occur downstream from generation of reactive oxygen species occurring independently of wt p53 during X-irradiation-induced cell death.

Mutational analysis of the beta-catenin gene in gastric carcinomas

Y Sasaki, Morimoto, I, M Kusano, M Hosokawa, F Itoh, K Yanagihara, K Imai, T Tokino

TUMOR BIOLOGY ( KARGER ) 22 ( 2 ) 123 - 130 2001.03

Previous studies reported that mutation of the adenomatous polyposis coli (APC) gene was not observed in the majority of gastric cancers. To evaluate the role of the APC/beta -catenin/Tcf pathway, we analyzed mutations in the beta -catenin gene and the accumulation of beta -catenin protein in gastric carcinomas. An interstitial deletion spanning exon 3 of the beta -catenin gene was observed in 1 of 13 gastric cancer cell lines. No missense mutation was found in these 13 cell lines. Nuclear and/or cytoplasmic localization of beta -catenin was observed in 16 of 70 primary gastric carcinomas by immunohistochemistry, while we found no mutations in exon 3 in 35 carcinoma tissues available for PCR amplification. Our findings suggest that somatic mutations of the beta -catenin gene are rare in human gastric carcinomas and that accumulation of normal beta -catenin protein in a subset of gastric cancers may be due to other mechanisms of its activation. Copyright (C) 2001 S. Karger AG, Basel.

アデノウイルスベクターを用いたp53ファミリー遺伝子導入は大腸癌由来細胞株に細胞周期停止とアポトーシスを誘導する一大腸癌の遺伝子治療への応用

佐々木泰史, 時野隆至

放射線生物研究 36 ( 3 ) 2001

潰よう性大腸炎におけるメチル化の異常と発癌リスク予測への応用

豊田実, 伊東文生, 菊地剛史, 鈴木拓, 小畑俊郎, 佐藤亜由美, 佐々木泰史, 時野隆至, 今井浩三

日本癌学会総会記事 60th 2001

p53標的遺伝子としてのオステオポンチン遺伝子の同定

森本一郎, 佐々木泰史, 石田勢津子, 伊東文生, 今井浩三, 時野隆至

日本癌学会総会記事 60th 2001

p53ノックアウトマウスを用いたp53標的遺伝子の同定

石田勢津子, 豊田実, 佐々木泰史, 森本一郎, 伊東文生, 今井浩三, 時野隆至

日本癌学会総会記事 60th 2001

ネットワーク・マルチパラレル・コンピューティングによるp53結合配列解析システムの開発

明石浩史, 豊田実, 青木文夫, 佐々木泰史, 宮司正道, 西森博幸, 今井浩三, 時野隆至, 辰巳治之

日本癌学会総会記事 60th 2001

p53類似遺伝子(p73,p51)を用いた大腸癌に対する遺伝子治療の基礎的研究

佐々木泰史, 坂本裕史, 時野隆至

日本消化器病学会雑誌 98 2001

p53類似遺伝子p73,p63/p51の標的遺伝子の検索

佐々木泰史, 森本一郎, 石田勢津子, 今井浩三, 鬼原史, 田中敏博, 中村祐輔, 時野隆至

日本癌学会総会記事 60th 2001

p53AlP1, a potential mediator of p53-dependent apoptosis, and its regulation by Ser-46-phosphorylated p53

K Oda, H Arakawa, T Tanaka, K Matsuda, C Tanikawa, T Mori, H Nishimori, K Tamai, T Tokino, Y Nakamura, Y Taya

CELL ( CELL PRESS ) 102 ( 6 ) 849 - 862 2000.09

Through direct cloning of p53 binding sequences from human genomic DNA, we have isolated a novel gene, designated p53AIP1 (p89-regulated Apoptosis-Inducing Protein 1), whose expression is inducible by wildtype p53. Ectopically expressed p53AIP1, which is localized within mitochondria, leads to apoptotic cell death through dissipation of mitochondrial Delta psi m. We have found that upon severe DNA damage, Ser-46 on p53 is phosphorylated and apoptosis is induced. In addition, substitution of Ser-46 inhibits the ability of p53 to induce apoptosis and selectively blocks expression of p53AIP1. Our results suggest that p53AIP1 is likely to play an important role in mediating p53-dependent apoptosis, and phosphorylation of Ser-46 regulates the transcriptional activation of this apoptosis-inducing gene.

子宮内膜癌におけるSRCの変異

小林寛治, 寒河江悟, 杉村政樹, 西岡善宏, 時野隆至, 工藤隆一

日本癌学会総会記事 ( 日本癌学会 ) 59回 565 - 566 2000.09

A single nucleotide polymorphism in the matrix metalloproteinase-1 promoter in endometrial carcinomas

Y Nishioka, K Kobayashi, S Sagae, S Ishioka, A Nishikawa, M Matsushima, Y Kanamori, T Minaguchi, Y Nakamura, Takashi Tokino, R Kudo

JAPANESE JOURNAL OF CANCER RESEARCH ( BUSINESS CENTER ACADEMIC SOCIETIES JAPAN ) 91 ( 6 ) 612 - 615 2000.06

Recent studies demonstrated that a single guanine insertion polymorphism in a matrix metallopretease-1 promoter created an Ets binding site and affected the elevation of the transcriptional level of matrix metalloproteinase-1 (MMP-1). Furthermore, in tumor cell lines derived from melanoma and breast cancer, the incidence of the 2G/2G genotype was significantly higher than that in the normal population, To evaluate the contribution of this polymorphism in endometrial carcinomas, we genotyped 100 endometrial carcinomas and then analyzed immunoexpression of MMP-1 in these carcinomas. We found that endometrial carcinoma patients showed a significantly higher rate of 1G/2G or 2G/2G genotype than control individuals, and that tumors containing the 2G allele(a) expressed MMP-1 protein more frequently than those,vith 1G/1G genotype. Therefore, the single nucleotide polymorphism at the MMP-1 promoter affected the expression level of the MMP-1 protein, which may result in the association with more aggressive character in endometrial carcinoma. Our result suggests that the presence of 2G polymorphism at the, MMP-1 promoter may be one of the risk factors for the development and/or progression of endometrial carcinoma.

Induction of cellular genes is mediated by the Bel1 transactivator in foamy virus-infected human cells

A Wagner, A Doerks, M Aboud, A Alonso, T Tokino, RM Flugel, M Lochelt

JOURNAL OF VIROLOGY ( AMER SOC MICROBIOLOGY ) 74 ( 10 ) 4441 - 4447 2000.05

To gain insight into human foamy virus (HFV; also called spumaretrovirus)-induced alterations of cellular genes, the expression profiles of defined genes in HFV-infected primary human cells were analyzed by cDNA array assays. Several distinct cellular genes activated by HFV infection were identified; the identities of the cellular genes were confirmed by RNA blot analyses. Compared with mock-infected controls, the concentrations of cellular Kip2, Egr-1, COUP-TF1, insulin-like growth factor II (IGF-II), and EphB3 mRNAs were significantly increased in HFV-infected cells and showed a gene-specific and time-dependent induction. Immunoblot analyses with antibodies against some of the cellular gene products revealed increased levels of the corresponding proteins. To investigate mechanisms of HFV-induced alterations in cellular gene expression, the capacity of known HFV genes to increase expression of defined cellular genes was analyzed by transient expression experiments. Plasmids that encode the HFV Bell transcriptional transactivator were necessary and sufficient to strongly increase expression of p57Kip2, IGF-II, and EphB3 genes in 293T cells. Potential mechanisms and consequences of activation of cellular genes during HFV infection and Bell transactivation of the Kip2 gene are discussed.

Nora, a BH3-only member of the Bcl-2 family and candidate mediator of p53-induced apoptosis

E Oda, R Ohki, H Murasawa, J Nemoto, T Shibue, T Yamashita, T Tokino, T Taniguchi, N Tanaka

SCIENCE ( AMER ASSOC ADVANCEMENT SCIENCE ) 288 ( 5468 ) 1053 - 1058 2000.05

A critical function of tumor suppressor p53 is the induction of apoptosis in cells exposed to noxious stresses. We report a previously unidentified pro-apoptotic gene, Nora. Expression of Nora induction in primary mouse cells exposed to x-ray irradiation was dependent on p53, Nora encodes a Bcl-2 homology 3 (BH3)-only member of the Bcl-2 family of proteins; this member contains the BH3 region but not other BH domains. When ectopically expressed, Nora underwent BH3 motif-dependent Localization to mitochondria and interacted with anti-apoptotic Bcl-2 family members, resulting in the activation of caspase-9. We also demonstrate that blocking the endogenous Nora induction results in the suppression of apoptosis. Nora may thus represent a mediator of p53-dependent apoptosis.

Isolation and characterization of a novel TP53-inducible gene, TP53TG5, which suppresses growth and shows cell cycle-dependent transition of expression

S Isaka, Y Takei, T Tokino, K Koyama, Y Miyoshi, M Suzuki, E Takahashi, C Azuma, Y Murata, Y Nakamura

GENES CHROMOSOMES & CANCER ( WILEY-LISS ) 27 ( 4 ) 345 - 352 2000.04

Through a strategy of direct cloning of TP53-binding DNA sequences from human genome DNA, we have identified a novel TP53-target gene, termed TP53TG5 (TP53-target gene 5). This gene, localized to chromosome band 20q13.1 by fluorescence in situ hybridization, encodes a 290-amino-acid peptide with no significant homology with any known proteins in the public database. A colony-formation assay using human glioblastoma cell line T98G, which lacks wild-type TP53 and expresses no endogenous TP53TG5, revealed a growth-suppressive effect of the TP53TG5 gene product. Furthermore, immunohistochemical studies, following transfection of T98G with plasmid designed to express green fluorescent protein-fused TP53TG5, revealed cell cycle-dependent intracellular localization of this protein. Our results suggest that functional studies of TP53TG5 may provide new insights into the complex physiological activities of TP53. (C) 2000 Wiley-Liss, Inc.

Mutation of the SRC gene in endometrial carcinoma

M Sugimura, K Kobayashi, S Sagae, Y Nishioka, S Ishioka, K Terasawa, T Tokino, R Kudo

JAPANESE JOURNAL OF CANCER RESEARCH ( BUSINESS CENTER ACADEMIC SOCIETIES JAPAN ) 91 ( 4 ) 395 - 398 2000.04

Recently, an activating mutation of the SRC gene has been implicated in about one-tenth of advanced colon cancers. The SRC 531 mutation results in truncation of SRC directly C-terminal to the regulatory Tyr 530 and appears to activate the Tyr 530, To investigate whether mutation of SRC plays an important role in the development and progression of gynecological tumors, we performed mutational analysis of the entire coding region of SRC in 70 ovarian carcinomas, 68 endometrial carcinomas and 3 endometrial stromal sarcomas by means of polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) followed by nucleotide sequencing and restriction fragment Length polymorphism (RFLP) analysis. We found one truncated mutation at codon 531 (Gln to Stop) in an endometrial carcinoma. However, we found no mutation of this gene in ovarian carcinoma or endometrial stromal sarcoma. Our results suggest that mutation of SRC may be implicated in a small proportion of endometrial carcinomas.

AXIN1 mutations in hepatocellular carcinomas, and growth suppression in cancer cells by virus-mediated transfer of AXIN1

S Satoh, Y Daigo, Y Furukawa, T Kato, N Miwa, T Nishiwaki, T Kawasoe, H Ishiguro, M Fujita, T Tokino, Y Sasaki, S Imaoka, M Murata, T Shimano, Y Yamaoka, Y Nakamura

NATURE GENETICS ( NATURE AMERICA INC ) 24 ( 3 ) 245 - 250 2000.03

The Wnt signalling pathway is essential for development and organogenesis(1-3). Wnt signalling stabilizes beta-catenin, which accumulates in the cytoplasm, binds to T-cell factor (TCF; also known as lymphocyte enhancer-binding factor, LEF) and then upregulates downstream genes(4-6). Mutations in CTNNB1 (encoding beta-catenin) or ape (adenomatous polyposis coli) have been reported in human neoplasms including colon cancers and hepatocellular carcinomas(7-13) (HCCs), Because HCCs tend to show accumulation of beta-catenin more often than mutations in CTNNB1, we looked for mutations in AXIN1, encoding a key factor for Wnt signalling, in 6 HCC cell lines and 100 primary HCCs. Among the 4 cell lines and 87 HCCs in which we did not detect CTNNB1 mutations, we identified AXIN1 mutations in 3 cell lines and 6 mutations in 5 of the primary HCCs. In cell lines containing mutations in either gene, we observed increased DNA binding of TCF associated with beta-catenin in nuclei. Adenovirus mediated gene transfer of wild-type AXIN1 induced apoptosis in hepatocellular and colorectal cancer cells that had accumulated beta-catenin as a consequence of either APC, CTNNB1 or AXIN1 mutation, suggesting that axin may be an effective therapeutic molecule for suppressing growth of hepatocellular and colorectal cancers.

Adenovirus-mediated transfer of p53-related genes induces apoptosis of human cancer cells

S Ishida, T Yamashita, U Nakaya, T Tokino

JAPANESE JOURNAL OF CANCER RESEARCH ( BUSINESS CENTER ACADEMIC SOCIETIES JAPAN ) 91 ( 2 ) 174 - 180 2000.02

Two p53-related genes, p73 and p51, were recently identified as structural homologues of the p53 tumor suppressor gene, suggesting that the roles of these tno genes may be similar to those of p53, including growth suppression and induction of apoptosis, Here we show that introduction of p73 or p51 cDNAs into cultured human cancer cells suppressed colony formation in the presence of G418, We then examined the ability of various isoforms of p73 and p51 to activate transcription of a reporter gene. This assay showed that p73 beta and p51A activated transcription through a consensus p53 binding sequence, while p73 alpha and p51B isoforms minimally transactivated the p53 reporter gene. To characterize further the biological functions of the p53-related genes, we constructed recombinant adenoviruses containing the p73 and p51 cDNAs. Ad-p73 beta and Ad-pS1A induced endogenous p21 gene expression more effectively than Ad-p73 alpha and Ad-p51B, respectively. To evaluate the mode of cell death induced by p53-related genes, Ad-p73 and Ad-D51 were used to infect human cancer cells. Infection of Ad-p73 beta, Ad-pS1A or Ad-p51B resulted in DNA fragmentation in a subset of cancer cell lines more efficiently than did infection of Ad-p53. We then examined the combined effect of each p53-related gene and the EIA oncogene in the induction of apoptosis, The EIA oncogene cooperated with p51 as well as p53 to induce apoptosis, while p73 resulted in a weak induction of apoptosis by EIA. Overall, apoptosis induction by p51B and p73 alpha isoforms may he due to mechanisms other than transcriptional activation of p53-target genes. Our results suggest that p53-related genes are both similar to and different from p53 in their pathways leading to growth suppression.

The role of p53-target genes in human cancer

Takashi Tokino, Y Nakamura

CRITICAL REVIEWS IN ONCOLOGY HEMATOLOGY ( ELSEVIER SCIENCE INC ) 33 ( 1 ) 1 - 6 2000.01

Book review, literature introduction, etc.

The tumor suppressor gene p53 is mutated in a large proportion of human cancers. In some cellular conditions like DNA damage, the p53 gene is induced and its gene product is posttranscriptionally activated. p53 works as a transcriptional activator and induces the expression of its downstream target genes. This review will explain why expression of the normal p53 gene leads to tumor growth suppression. The p53 has several biological effects involving cell-cycle arrest, DNA replication and repair, proliferation, apoptosis, angiogenesis inhibition, and cellular stress response. These effects of the p53 result mainly from the activation of expression of a large number of p53-target genes. Here we have focused on the biological functions of the transcriptional targets of p53. (C) 2000 Elsevier Science Ireland Ltd. All rights reserved.

p53類似遺伝子p73,p51の抗腫よう効果についての検討

佐々木泰史, 森本一郎, 中谷宇一郎, 石田勢津子, 今井浩三, 鬼原史, 田中敏博, 中村祐輔, 時野隆至

Japanese Journal of Cancer Research 91 ( Supplement (Sept) ) 2000

Ewing肉腫特異的なEWS-ETS融合転写因子の標的遺伝子

吉田幸一, 佐々木泰史, 梅沢明弘, 秦順一, 鬼原史, 田中敏博, 中村祐輔, 時野隆至

Japanese Journal of Cancer Research 91 ( Supplement (Sept) ) 2000

頭頚部癌におけるβ-catenin遺伝子の蛋白発現とmutation,増殖因子受容体の発現

小田島哲世, 佐々木泰史, 山口晃, 田中信幸, 池田健, 佐藤昌明, 時野隆至

Japanese Journal of Cancer Research 91 ( Supplement (Sept) ) 2000

p53ノックアストマウスを用いたp53標的遺伝子の同定

森本一郎, 佐々木泰史, 石田勢津子, 中谷宇一郎, 山下利春, 伊東文生, 今井浩三, 時野隆至

Japanese Journal of Cancer Research 91 ( Supplement (Sept) ) 2000

Ewing肉腫/PNETに特異的EWS-ETS融合転写因子の標的遺伝子

吉田幸一, 佐々木泰史, 梅沢明弘, 秦順一, 鬼原史, 田中敏博, 中村祐輔, 時野隆至

DNAチップ技術研究会プログラム・抄録集 1st 2000

ヒト大腸癌由来細胞株におけるp53類似遺伝子p73,p51によるアポトーシス誘導と抗癌剤との併用効果

石田勢津子, 佐々木泰史, 森本一郎, 中谷宇一郎, 山下利春, 今井浩三, 時野隆至

Japanese Journal of Cancer Research 91 ( Supplement (Sept) ) 2000

Localization of membrane-associated guanylate kinase (MAGI)-1/BAI-associated protein (BAP) 1 at tight junctions of epithelial cells

N Ide, Y Hata, H Nishioka, K Hirao, Yao, I, M Deguchi, A Mizoguchi, H Nishimori, T Tokino, Y Nakamura, Y Takai

ONCOGENE ( STOCKTON PRESS ) 18 ( 54 ) 7810 - 7815 1999.12

Membrane-associated guanylate kinase (MAGI)-1/BAI-associated protein (BAP) 1 and Synapse-associated protein (SAP) 97/human Discs-large tumor suppressor gene (hDLG) are ubiquitous isoforms of synaptic scaffolding molecule (S-SCAM) and Postsynaptic density(PSD)-95/SAP90, both of which are implicated in the structures of synapses, respectively. SAP97/hDLG is localized at epithelial junctions and may function as a scaffolding protein, but the subcellular localization or the function of MAGI-1/BAP1 has not been clarified. In intestinal epithelial cells, MAGI-1/BAP1 was localized at tight junctions, whereas SAP97/hDLG was localized diffusely at cell-cell junctions. In Madine Darby canine kidney (MDCK) cells, MAGI-1/BAP1 was colocalized with ZO-1, whereas SAP97/hDLG was colocalized with E-cadherin. In MDCK cells, dominant active and negative mutants of Rad small G protein changed the amounts of SAP97/hDLG at cell-cell junctions, but not that of MAGI-1/BAP1. When MDCK cells were switched to a low Ca2+ medium, E-cadherin disappeared from the plasma membrane, and cells were dissociated. The phorbol 12-myristate 13-acetate-treatment after the low Ca2+ switch induced a tight junction-like structure. MAGI-1/BAP1 was recruited with ZO-1 to this structure, but SAP97/hDLG or E-cadherin was not. These findings suggest that MAGI-1/BAP1 is a component of tight junctions of epithelial cells, and that its role is different from that of SAP97/hDLG.

Induction of transformation and p53-dependent apoptosis by adenovirus type 5 E4orf6/7 cDNA

S Yamano, T Tokino, M Yasuda, M Kaneuchi, M Takahashi, Y Niitsu, K Fujinaga, T Yamashita

JOURNAL OF VIROLOGY ( AMER SOC MICROBIOLOGY ) 73 ( 12 ) 10095 - 10103 1999.12

Adenovirus (Ad) E4orf6/7, one of the early gene products of human Ads, forms a stable complex with the cellular transcription factor E2F to activate transcription from the Ad E2 promoter. E2F cDNAs have growth-promoting and apoptosis-inducing activities when overexpressed in cells. We cloned Ad5 E40rf6/7 cDNA in both simian virus 40- and human cytomegalovirus-based expression vectors to examine its transforming and apoptotic activities. The cloned E4orf6/7 collaborated with a retinoblastoma protein (RB)-nonbinding and therefore E2F-nonreleasing mutant of Ad5 ELA (dl922/947) to morphologically transform primary rat cells, suggesting that E2F is an important cellular protein functioning downstream of E1A for transformation. In a G418 colony formation assay, E4orf6/7 was shown to suppress growth of untransformed rat cells. Moreover, a recombinant Ad expressing Ad5 E4orf6/7 induced apoptosis in rat cells when coinfected with wild-type p53-expressing Ad. Mutational analysis of E4orf6/7 revealed that both of the domains required for growth inhibition and transformation by E4orf6/7 lay in the C-terminal region, which is essential for transactivation from the upstream sequence of an E2a promoter containing E2F-binding sites. However, the smallest mutant of E4orf6/7, encoding the C-terminal 59 amino acids, failed to complement the RE-nonbinding dl/922/947 mutant despite showing growth inhibition and E2F transactivation activities. Thus, it is suggested that a subregion of E40rf6/7 which is required for growth inhibition and transformation in collaboration with dl922/947 overlaps the transactivation domain of E4orf6/7.

Retrovirus vectors designed for efficient transduction of cytotoxic or cytostatic genes

M Ui, M Takada, T Arai, K Matsumoto, K Yamada, T Nakahata, T Nishiwaki, Y Furukawa, Takashi Tokino, Y Nakamura, H Iba

GENE THERAPY ( STOCKTON PRESS ) 6 ( 10 ) 1670 - 1678 1999.10

It is difficult to establish stable packaging cell lines producing retrovirus vectors for the expression of anti-oncogenes with cytotoxic or cytostatic potential, because these genes would also affect the growth of the packaging cell lines. To overcome this problem, we designed a transcriptional unit pBabeLPL for vector RNA production, in which the transcription of the exogenous genes is completely suppressed by the presence of a preceding insertion containing the puromycin resistance gene (puro) and a poly(A) addition signal. This insertion is flanked by a tandem pair of loxP, and is designed to be excised after the introduction of Cre recombinase, when transcription of the exogenous gene will be started from the 5'-LTR. The transcriptional unit carrying LacZ or p53 as the exogenous gene was introduced into a previously constructed prepackaging cell lines PtG-S2, in which the expression of VSV-G is also designed to be initiated by the introduction of Cre recombinase, while the gag-pol gene is expressed continuously. After the introduction of Cre recombinase by an adenovirus vector, LacZ- or p53- expressing VSV-G-pseudotyped retrovirus vectors with the designed structure were produced at high virus titers. The p53 virus was shown to be able to transduce p53 into the entire population of several human cancer cell lines and to induce their growth arrest at the GI phase, indicating that this vector-producing system will be advantageous for human gene therapy.

細胞傷害性や細胞増殖抑制活性を持つ遺伝子発現用レトロウイルスベクターの設計と開発

宇井基泰, 新井徹, 中畑龍俊, 西脇忠, 古川洋一, 時野隆至, 中村祐輔, 伊庭英夫

日本癌学会総会記事 ( 日本癌学会 ) 58回 319 - 319 1999.08

Mutational analysis of STK11 gene in ovarian carcinomas

Y Nishioka, K Kobayashi, S Sagae, M Sugimura, S Ishioka, M Nagata, K Terasawa, T Tokino, R Kudo

JAPANESE JOURNAL OF CANCER RESEARCH ( ELSEVIER SCI IRELAND LTD ) 90 ( 6 ) 629 - 632 1999.06

Recently STK11, the causative gene of Peutz-Jeghers syndrome (PJS) was identified on chromosome 19p13.3. PJS is often accompanied by several malignancies, including breast tumor, adenoma malignum of the uterine cervix, and ovarian tumor. To investigate the involvement of STK11 gene in the development of ovarian carcinomas, we analyzed 30 ovarian carcinomas for loss of heterozygosity (LOH) and STK11 gene mutations. We found one missense mutation (codon 281, Pro to Leu) with heterozygous and somatic status. This mutation occurred at codon 281, which lies within the mutational hot spot (codon 279-281) of STK11 gene previously reported in PJS. We also detected LOH in 2 (11%) of 19 informative ovarian carcinomas. Our results suggest that mutations of the STK11 gene may play a limited role in the development of ovarian carcinomas.

Mutational analysis of beta-catenin gene in Japanese ovarian carcinomas: Frequent mutations in endometrioid carcinomas

S Sagae, K Kobayashi, Y Nishioka, M Sugimura, S Ishioka, M Nagata, K Terasawa, T Tokino, R Kudo

JAPANESE JOURNAL OF CANCER RESEARCH ( ELSEVIER SCI IRELAND LTD ) 90 ( 5 ) 510 - 515 1999.05

To investigate the contribution of the beta-catenin gene to the development of ovarian carcinomas, mutational analysis of exon 3 of the beta-catenin gene was conducted. We analyzed 61 primary ovarian carcinomas, consisting of 49 non-endometrioid-type and 12 endometrioid-type tumors, for genetic alteration of the beta-catenin gene, Five carcinomas showed beta-catenin mutations (S37C, T41I, T41A), including 4 (33%) of 12 endometrioid-type tumors and 1 (14%) of 7 mucinous-type tumors. All of these mutations altered at the serine/threonine residues that are potential sites of GSK3-beta phosphorylation, We detected no carcinomas with interstitial deletion involving exon 3 of beta-catenin, Furthermore, we immunohistochemically studied 27 of the 61 ovarian carcinomas. Both nuclear and cytoplasmic beta-catenin expressions were demonstrated in 3 of the 27 ovarian carcinomas for which tissue samples were available for examination. All 4 cases exhibited mutations in exon 3 of beta-catenin, including a mucinous carcinoma. Our results suggested that beta-catenin gene mutation at potential GSK3-beta phosphorylation sites results in accumulation of beta-catenin protein within the cells and its translocation to nuclei. Accumulated beta-catenin protein may be involved in the development of endometrioid-type ovarian carcinomas, and some mucinous-type ovarian carcinomas.

Recombinant adenovirus expressing wild-type p53 is antiangiogenic: A proposed mechanism for bystander effect

M Nishizaki, T Fujiwara, T Tanida, A Hizuta, H Nishimori, Takashi Tokino, Y Nakamura, M Bouvet, JA Roth, N Tanaka

CLINICAL CANCER RESEARCH ( AMER ASSOC CANCER RESEARCH ) 5 ( 5 ) 1015 - 1023 1999.05

Angiogenesis is required for the growth and progression of malignancies, Recent studies have demonstrated that genetic alterations may accompany acquisition of the angiogenic phenotype, The tumor suppressor gene p53 is most frequently mutated in human cancers and is also known to be a transcriptional regulator of a variety of genes. Here, we investigated the antiangiogenic effect of the wild-type p53 (wt-p53) gene transfer on a human non-small cell lung cancer cell line. Mutant p53-expressing H226Br non-small cell lung cancer cells were transduced with the wt-p53 gene using a recombinant adenoviral vector (Ad5CMVp53) and applied to semiquantitative reverse transcription-PCRs for the detection of altered mRNA expression of angiogenic and/or antiangiogenic factors. In vivo neovascularization assay of Ad5CMVp53-infected cells was then performed using a membrane-diffusion chamber system s.c. transplanted in nu/nu mice. We also evaluated the effect of Ad5CMVp53-infected H226Br cells on nontransduced tumor cells in vivo by s.c. inoculating mixture of cells into nu/nu mice. Ad5CMVp53 infection markedly inhibited the expression of an angiogenic factor, vascular endothelial growth factor, and increased the expression of a novel antiangiogenic factor, brain-specific angiogenesis inhibitor 1, resulting in reduced neovascularization in vivo. Mixing experiments showed that tumor cells transduced with the wt-p53 gene inhibited the in vivo tumor growth of adjacent nontransduced cells. Our data suggest that a recombinant adenovirus expressing the wt-p53 gene is antiangiogenic, which may explain, in part, the mechanism of the bystander effect induced by the wt-p53 gene transfer on adjacent tumor cells.

Mutations of the beta-catenin gene in endometrial carcinomas

K Kobayashi, S Sagae, Y Nishioka, T Tokino, R Kudo

JAPANESE JOURNAL OF CANCER RESEARCH ( ELSEVIER SCI IRELAND LTD ) 90 ( 1 ) 55 - 59 1999.01

To investigate the contribution of beta-catenin to the development of endometrial carcinoma, we searched for genetic alterations of the beta-catenin gene in primary endometrial carcinomas. Mutational analysis of exon 3 of the beta-catenin gene, encoding the serine/threonine residues for GSK-3 beta phosphorylation, was performed for 35 tumors. Nucleotide sequencing analysis revealed that 5 tumors (5/35, 14%) contained mutations (S33C, S37C, S37F, T41A) that altered potential GSK-3 beta phosphorylation sites. Each of the mutations resulted in the substitution of serine/threonine residues that have been implicated in the down-regulation of beta-catenin through phosphorylation by GSK-3 beta kinase, Furthermore, the incidence of beta-catenin mutations was significantly higher in early-onset (3 of 5) than that in late-onset tumors (2 of 30) (P = 0.014, Fisher's exact test). Replication error (RER)-positive phenotype was not detected in tumors with the beta-catenin gene mutation, although 10 of 35 tumors revealed RER, We performed immunohistochemistry of beta-catenin in 17 cases for which tissue samples were available. We confirmed accumulation of beta-catenin protein in both the nucleus and cytoplasm in 3 tumors, including two in which amino acid alterations had occurred at codon 33 and 37, The other case had no mutation in exon 3, Our results suggested that mutations at serine/threonine residues involved in phosphorylation by GSK-3 beta affected the stability of beta-catenin, Accumulation of mutant beta-catenin could contribute to the development of a subset of endometrial carcinomas, particularly those of the early-onset type.

Molecular cloning, mapping, and characterization of a novel human gene, MTA1-L1, showing homology to a metastasis-associated gene, MTA1

M Futamura, H Nishimori, T Shiratsuchi, S Saji, Y Nakamura, T Tokino

JOURNAL OF HUMAN GENETICS ( SPRINGER-VERLAG TOKYO ) 44 ( 1 ) 52 - 56 1999

Through large-scale sequencing of clones randomly selected from libraries of human cDNAs, we have isolated a novel human gene encoding a product with 59.6% identity in amino acid sequence to human MTA1, a protein associated with tumor invasion and metastasis. This cDNA, named MTA1-L1 (MTA1 like 1), consists of 2736 nucleotides with an open reading frame encoding 668 amino acids. A single 3.0-kb transcript of MTA1-L1 was expressed ubiquitously on Northern blots. Structural analysis of the MTA1-L1 gene revealed 18 exons spanning 8.1 kb of genomic DNA. We assigned the MTA1-L1 locus to chromosomal band 11q12-13.1 by fluorescence in situ hybridization.

P-2 子宮内膜癌におけるβ-catenin遺伝子の変異

Induction of apoptosis in T98G glioblastoma cells by transfection of GML, a p53 target gene

K Ueda, Y Miyoshi, T Tokino, M Watatani, Y Nakamura

ONCOLOGY RESEARCH ( COGNIZANT COMMUNICATION CORP ) 11 ( 3 ) 125 - 132 1999

Expression of the glycosyl-phosphatidylinositol-anchored molecule-like protein (GML) gene, a p53 target, correlates with the sensitivity of some cancer cell lines to anticancer drugs and ionizing radiation. To investigate the function of GML further, we introduced the GML cDNA into various cancer cell lines under control of the tetracycline-regulated system. When we introduced GML into human glioblastoma cell line T98G, which lacks wild-type p53 and expresses no endogenous GML, we observed significant growth suppression accompanied by G(2)/M arrest in two independent, stable cell lines. We confirmed induction of apoptosis by fluorescence-activated cell sorting (FACS) analysis and nuclear staining. Our results indicated that GML could induce apoptosis of T98G without functional p53, and implied that GML plays a crucial role in the apoptotic pathway in some cancer cells.

Significant increase of adenovirus infectivity in glioma cell lines by extracellular domain of hCAR

T Mori, H Arakawa, T Tokino, K Mineura, Y Nakamura

ONCOLOGY RESEARCH ( COGNIZANT COMMUNICATION CORP ) 11 ( 11-12 ) 513 - 521 1999

Recombinant adenoviruses are highly advantageous as vectors for transferring genes into mammalian cells, but the transfer is not efficient in all types of cells. We investigated the effects of four adenoviral receptors [integrin alpha v, integrin beta 3, integrin beta 5, and human coxsackievirus and adenovirus receptor (hCAR)] on adenovirus-mediated transfer of exogenous cDNA into each of 10 glioma cell lines. Transfection efficiency varied widely from one cell line to another (0-100%) when we measured it by infection with AdLacZ, a vector designed to express P-galactosidase. Levels of integrin alpha v and integrin beta 5 expression were similar among the 10 cell lines, but expression of hCAR and integrin beta 3 varied significantly. As these observations indicated a possible correlation between expression of hCAR and the efficiency of gene transfer, we induced the hCAR gene into three glioma cell lines (T98G, U118MG, and U138MG) that expressed hCAR at very low levels and had also revealed low efficiencies of adenoviral gene transfer. In U118MG- and U138MG-derived cells that had regained the ability to express hCAR in stable fashion, adenovirus-mediated gene transfer became highly efficient. Moreover, addition of the peptide corresponding to the extracellular domain of hCAR (ECD-hCAR) by preincubation significantly increased the adenovirus infectivity to these adenovirus-tolerant cells. These results suggest that hCAR could be one of important determinants of the infectivity of adenovirus: and that the ECD-hCAR might be a novel useful tool for improvement of adenovirus-mediated gene therapy against the adenovirus-tolerant cancer cells.

Identification of BAIAP2 (BAI-associated protein 2), a novel human homologue of hamster IRSp53, whose SH3 domain interacts with the cytoplasmic domain of BAI1

K Oda, T Shiratsuchi, H Nishimori, J Inazawa, H Yoshikawa, Y Taketani, Y Nakamura, T Tokino

CYTOGENETICS AND CELL GENETICS ( KARGER ) 84 ( 1-2 ) 75 - 82 1999

BAI1 (brain-specific angiogenesis inhibitor 1) was originally isolated as a p53-target gene specifically expressed in brain. To clarify its function, we have been searching for cellular proteins that associate with the cytoplasmic domain of BAI1. Using its intracellular carboxyl terminus as "bait" in a yeast two-hybrid system, we isolated a cDNA clone named BAIAP2 whose nucleotide sequence would encode a 521-amino acid protein showing significant homology to a 58/53-kDa substrate of insulin-receptor kinase in the hamster. As the expression profile of BAIAP2 examined by Northern blot analysis was almost identical to that of BAI 1, BAIAP2 appears to be active mainly in neurons. In vitro binding assays confirmed that a proline-rich cytoplasmic fragment of BAI1 interacted with the Src homology 3 (SH3) domain of BAIAP2. Double-color immunofluorescent analysis revealed that BAIAP2 was localized at the cytoplasmic membrane when it was coexpressed with BAI1 in COS-7 cells; BAIAP2 not associated with BAI1 was diffused in the cytoplasm. Predominant localization of BAI1 protein in a sub-cellular fraction enriched in growth cones indicated a possible role of BAI1 as a cell adhesion molecule inducing growth cone guidance. As a protein partner of BAI1, BAIAP2 may represent an important link between membrane and cytoskeleton in the process of neuronal growth.

小林寛治, 寒河江悟, 西岡嘉宏, 時野隆至, 工藤隆一

日本産科婦人科學會雜誌 ( 日本産科婦人科学会 ) 51 "S - 360" 1999

胃癌におけるβ-カテニンの発現性と遺伝子異常の検索

佐々木泰史, 森本一郎, 伊東文生, 柳原五吉, 今井浩三, 時野隆至

日本癌学会総会記事 58th 1999

Cloning and characterization of BAP3 (BAI-associated protein 3), a C2 domain-containing protein that interacts with BAI1

T Shiratsuchi, K Oda, H Nishimori, M Suzuki, E Takahashi, T Tokino, Y Nakamura

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS ( ACADEMIC PRESS INC ) 251 ( 1 ) 158 - 165 1998.10

BAI1 (brain-specific angiogenesis inhibitor 1), a p53-target gene specifically expressed in brain, encodes a seven-span transmembrane protein considered to be a member of the secretin receptor family. Using a two-hybrid system, we isolated a cDNA encoding a product that interacts with the cytoplasmic region of BAI1 and designated it BAP3 (BAI1-associated protein 3). The BAP3 product is a novel C2 domain-containing molecule with homology to Munc13 and synaptotagmin. As with Munc13, BAP3 is expressed predominantly in brain. Deletion-mutant analysis revealed that the interaction between BAI1 and BAP3 was not mediated by the Ca domains. Its predominant expression in brain and homology to Munc13 indicate that BAP3, by interacting with BAI1, might be involved in some neuronal function such as regulating release of neurotransmitters. (C) 1998 Academic Press.

Isolation of a novel TP53 target gene from a colon cancer cell line carrying a highly regulated wild-type TP53 expression system

Y Takei, S Ishikawa, T Tokino, T Muto, Y Nakamura

GENES CHROMOSOMES & CANCER ( WILEY-LISS ) 23 ( 1 ) 1 - 9 1998.09

We established a colon cancer cell line SW480-LOWTP53-I carrying a wild-type TP53 transgene that is inducible under control of the lactose operon. Induction of this transgene by isopropyl-beta-D-thiogalactoside (IPTG) arrests growth of the transfected cells. To investigate cellular responses related to the TP53 signaling pathway to induce growth arrest, we applied a differential display method to screen mRNAs isolated from this cell line and looked for genes whose expression was activated or suppressed after induction of wild-type TP53. Subsequent Northern blot analysis confirmed that expression of one novel gene was regulated by wild-type TP53. The cDNA, termed TP53TGI (TP53 target gene I), contained an open reading frame of 270 nucleotides encoding 90 amino acids. Under conditions of cellular stress (ultraviolet irradiation or exposure to bleomycin or cisplatin), expression of TP53TGI was induced in a wild-type TP53-dependent manner, indicating that this gene is likely to play an important role in the signaling pathway of TP53 and may function in response to cellular damage. (C) 1998 Wiley-Liss, Inc.

Cloning and characterization of BAI-associated protein 1: A PDZ domain-containing protein that interacts with BAI1

T Shiratsuchi, M Futamura, K Oda, H Nishimori, Y Nakamura, Takashi Tokino

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS ( ACADEMIC PRESS INC ) 247 ( 3 ) 597 - 604 1998.06

Brain-specific angiogenesis inhibitor 1 (BAI1), which is a p53-target gene specifically expressed in brain, encodes a seven-span transmembrane protein. Using a two-hybrid system, we isolated a cDNA that encodes a protein, named BAP1 (BAI1-associated protein), which interacts with the cytoplasmic region of BAI1. BAP1 is a novel member of the MAGUK (membrane-associated guanylate kinase homologue) family; it possesses a guanylate kinase domain, WW domains, and multiple PDZ domains. Interaction between BAI1 and BAP1 was mediated by a QTEV motifin the carboxy-terminal region of BAI1 and PDZ domains of BAP1, By immunocytochemical analysis of COS-7 cells transfected with BAI1 and BAP1, both products were co-localized at the cytoplasmic membrane, especially at cell-cell junctions. Cells transfected with BAI1 formed filopodia-like cytoplasmic extensions. These results suggest that BAI1 and BAP1 might be involved in cell adhesion and signal transduction in brain. (C) 1998 Academic Press.

CSR, a scavenger receptor-like protein with a protective role against cellular damage caused by UV irradiation and oxidative stress

HJ Han, Takashi Tokino, Y Nakamura

HUMAN MOLECULAR GENETICS ( OXFORD UNIV PRESS ) 7 ( 6 ) 1039 - 1046 1998.06

Oxidative stress is a pathogenic condition that causes cellular damage and, in a normally functioning cell, several transcription factors respond to this threat by modulating expression of genes whose products ameliorate the altered redox status in some way. We have isolated a novel macrophage scavenger receptorlike gene, CSR (cellular stress response), whose transcription in normal fibroblasts was significantly elevated by exposure to UV radiation or hydrogen peroxide, and pre-treatment with antioxidants prevented induction of CSR. Under conditions of oxidative stress, reactive oxygen species were significantly depleted in CSR-overexpressing cells, indicating that the CSR product protects cells by scavenging oxidative molecules or harmful products of oxidation. Further investigations into the regulation and function of CSR should open a way to understanding the cellular response and the pathogenic processes caused by oxidative stress.

Cloning and characterization of the murine P2XM receptor gene

G Nawa, T Urano, T Tokino, T Ochi, Y Miyoshi

JOURNAL OF HUMAN GENETICS ( SPRINGER-VERLAG TOKYO ) 43 ( 4 ) 262 - 267 1998

We have isolated the murine counterpart of the human P2XM gene (mP2XM), a P2X purinoceptor that is expressed predominantly in skeletal muscle. The mP2XM gene, consisting of 12 exons that span 10kb of genomic DNA, encodes a 379-amino-acid product with 83% identity to the human homologue. Two potential transmembrane domains (M1 and M2) are present in the predicted product, and a segment resembling the H5 region of voltage-gated ion channels is completely conserved between human and mouse P2XM proteins. Predominant expression of mP2XM in murine skeletal muscle was confirmed by Northern-blot analysis, and a low level of expression was detectable in lung tissue, although human lung does not express P2XM.

Overexpression of GML promotes radiation-induced cell cycle arrest and apoptosis

K Kagawa, T Inoue, T Tokino, Y Nakamura, T Akiyama

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS ( ACADEMIC PRESS INC ) 241 ( 2 ) 481 - 485 1997.12

Expression of the GML gene is regulated in a p53-dependent manner and is correlated with the sensitivity of esophageal cancer cells to anti-cancer drugs. To clarify the effect of GML expression on the sensitivity of cancer cells to ionizing radiation treatment, we established cell lines derived from p53-mutant human osteosarcoma HOS and esophageal carcinoma TE10 lines in which GML expression can be induced using the tetracycline-regulable system. Colony formation assay showed that the growth of cells expressing GML are inhibited in response to ionizing radiation, whereas cells not expressing GML were resistant to irradiation. Further investigation demonstrated that GML expression enhances G2/M arrest and apoptosis induced by gamma-irradiation. These results suggest that GML sensitizes cancer cells to ionizing radiation. (C) 1997 Academic Press.

A novel brain-specific p53-target gene, BAI1, containing thrombospondin type 1 repeats inhibits experimental angiogenesis

H Nishimori, T Shiratsuchi, T Urano, Y Kimura, K Kiyono, K Tatsumi, S Yoshida, M Ono, M Kuwano, Y Nakamura, T Tokino

ONCOGENE ( STOCKTON PRESS ) 15 ( 18 ) 2145 - 2150 1997.10

The genetic alteration of p53 is associated with neovascularization during progression of glioma to its more malignant form, glioblastoma, Hence, one or more of the genes transactivated by p53 is likely to function as an angiogenesis inhibitors, We isolated a novel p53-inducible gene that encodes a 1584-amino-acid product containing five thrombospondin type 1 (TSP-type 1) repeats and is specifically expressed in the brain, A recombinant protein corresponding to the TSP-type 1 repeats of this gene product inhibited in vivo neovascularization induced by bFGF in the rat cornea, The expression of this gene, designated BAI1 (brain-specific angiogenesis inhibitor 1) was absent or significantly reduced in eight of nine glioblastoma cell lines, suggesting BAI1 plays a significant role in angiogenesis inhibition, as a mediator of p53.

A novel brain-specific p53-target gene, BAI1, containing thrombospondin type1 repeats inhibits experimental angiogenesis.

T Tokino, H Nishimori, T Shiratsuchi, T Urano, Y Kimura, K Kiyono, K Tatsumi, S Yoshida, M Ono, M Kuwano, Y Nakamura

AMERICAN JOURNAL OF HUMAN GENETICS ( UNIV CHICAGO PRESS ) 61 ( 4 ) A84 - A84 1997.10

Research paper, summary (international conference)

GML sensitizes cancer cells to Taxol by induction of apoptosis

Y Kimura, T Furuhata, T Shiratsuchi, H Nishimori, K Hirata, Y Nakamura, T Tokino

ONCOGENE ( STOCKTON PRESS ) 15 ( 11 ) 1369 - 1374 1997.09

Recently we identified a novel gene, gml, whose expression is regulated in a p53-dependent manner and found that gml expression was correlated with the sensitivity of esophageal cancer cells to anticancer drugs. To further investigate the biological mechanism of gml in determining the chemosensitivity of cancer cells to clinically useful agents, we introduced gml cDNA into TE10, an esophageal cancer cell line that lacks endogenous gml expression. In two resulting stable cell lines which expressed gml cDNA in the absence of wild-type p53, cell death occurred within 6 h after treatment with Taxol. TE10 parent cells or TE10 cells transfected with vector alone displayed relative resistance for 36 h. Induction of gml did not by itself affect viability. Morphological analysis confirmed that the increased chemosensitivity to Taxol conferred by gml was due to apoptosis. These data suggest that reduced expression of gml is likely to be associated with poor response rates to chemotherapy, and that an assay for gml expression might serve a clinical purpose as a predictor of chemotherapeutic sensitivity.

Butyrate activates the WAF1/Cip1 gene promoter through Sp1 sites in a p53-negative human colon cancer cell line

K Nakano, T Mizuno, Y Sowa, T Orita, T Yoshino, Y Okuyama, T Fujita, N OhtaniFujita, Y Matsukawa, T Tokino, H Yamagishi, T Oka, H Nomura, T Sakai

JOURNAL OF BIOLOGICAL CHEMISTRY ( AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC ) 272 ( 35 ) 22199 - 22206 1997.08

Butyrate is a well known colonic luminal short chain fatty acid, which arrests cell growth and induces differentiation in various cell types. We examined the effect of butyrate on the expression of WAF1/Cip1, a potent inhibitor of cyclin-dependent kinases, and its relation to growth arrest in a p53-mutated human colon cancer cell line WiDr, Five millimolar butyrate completely inhibited the growth of WiDr and caused G(1)-phase arrest. WAF1/Cip1 mRNA was rapidly induced within 3 h by treatment with 5.0 mM butyrate, and drastic WAF1/Cip1 protein induction was detected. Using several mutant WAF1/Cip1 promoter fragments, we found that the butyrate responsive elements are two Spl sites at -82 and -69 relative to the transcription start site. We also found that a TATA element at -46 and two overlapping consensus Spl sites at -60 and -55 are essential for the basal promoter activity of WAF1/Cip1. These findings suggest that butyrate arrests the growth of WiDr by activating the WAF1/Cip1 promoter through specific Sp1 sites in a p53-independent fashion.

Cloning of P2XM a novel human P2X receptor gene regulated by p53

T Urano, H Nishimori, HJ Han, T Furuhata, Y Kimura, Y Nakamura, Takashi Tokino

CANCER RESEARCH ( AMER ASSOC CANCER RESEARCH ) 57 ( 15 ) 3281 - 3287 1997.08

Through cloning of functional p53-binding sites (p53-tagged sites) from the human genome, we isolated a novel gene inducible by wild-type p53, Its cDNA sequence contained an open reading frame encoding a 431-amino acid peptide that showed a significant homology with members of the P2X family, This protein also revealed a similarity to RP-2, a gene activated in thymocytes undergoing programmed cell death. Northern blot analysis showed that it was expressed predominantly in skeletal muscle, Hence, we designated the gene P2XM (P2X specifically expressed in skeletal muscle), P2XM was localized to chromosomal band 22q11, where frequent loss of heterozygosity has been observed in rhabdoid tumors, Although we detected no genetic alteration in the coding sequences, one of four rhabdomyosarcoma cell lines examined had completely lost expression of this gene, Furthermore, a minor splice variant lacking a part of exon 1 that would encode residues corresponding to transmembrane domain M1 was relatively more abundant in two of seven sarcoma cell lines, one of which was derived from a rhabdomyosarcoma, and the other was derived from an osteosarcoma. The results suggest that P2XM may play a significant role in the proliferation and/or differentiation of skeletal muscle cells and that its altered expression may be involved in the development of some sarcomas.

Genomic structure and chromosomal localization of GML (GPI-anchored molecule-like protein), a gene induced by p53

Y Kimura, T Furuhata, T Urano, K Hirata, Y Nakamura, T Tokino

GENOMICS ( ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS ) 41 ( 3 ) 477 - 480 1997.05

Among its known functions, tumor suppressor gene p53 serves as a transcriptional regulator and mediates various signals through activation of downstream genes, We recently identified a novel gene, GML (glycosylphosphatidylinositol (GPI)-anchored molecule-like protein), whose expression is specifically induced by wildtype p53, To characterize the GML gene further, we determined 35.8 kb of DNA sequence that included a consensus binding sequence for p53 and the entire GML gene. The GML gene consists of four exons, and the p53-binding sequence is present in the 5'-flanking region. in genomic organization this gene resembles genes encoding murine Ly-6 glycoproteins, a human homologue of the Ly-6 family called RIG-E, and CD59; products of these genes, known as GPI-anchored proteins, are variously involved in signal transduction, cell-cell adhesion, and cell-matrix attachment. FISH analysis revealed that the GML gene is located on human chromosome 8q24.3. Genes encoding at least two other GPI-anchored molecules, E48 and RIG-E, are also located in this region. (C) 1997 Academic Press.

Complete NotI restriction map covering the entire long arm of human chromosome 11

F Hosoda, Y Arai, E Kitamura, J Inazawa, M Fukushima, Takashi Tokino, Y Nakamura, C Jones, N Kakazu, T Abe, M Ohki

GENES TO CELLS ( BLACKWELL SCIENCE LTD ) 2 ( 5 ) 345 - 357 1997.05

Background: Human chromosome 11 is one of the autosomes on which many disease genes have been mapped. Many different types of map, including a radiation hybrid map, a genetic map, and an STS-content YAC map, have been constructed for the chromosome, However, a physical map providing accurate physical distances has not yet been established, A chromosome-wide NotI restriction map was constructed to understand the overall feature of the genome organization and to facilitate the positional cloning of disease genes. Results: A complete NotI restriction map of the entire long arm of human chromosome 11 was constructed using linking-clone mapping. This physical map covers 77.6 Mb, from a pericentromeric NotI site to the terminus, and provides the most accurate ordering and distance estimation to date. We also mapped 138 sequence markers in the q13 region that have been poorly mapped previously. Conclusions: The restriction map of the entire long arm of human chromosome 11 is the longest restriction map of the human genome. This mapping has disclosed unique features regarding the organization of the chromosome, indicating that restriction sites of NotI, a CpG-recognition enzyme, are primarily distributed in R (or T) bands and that genetic distance is considerably longer in R (or T) bands than in G bands. The mapping, as well as the dense concentration of mapped markers within the q13 region, should help with positional cloning of the genes associated with various diseases.

No evidence of mutation in the human PTC gene, responsible for nevoid basal cell carcinoma syndrome, in human primary squamous cell carcinomas of the esophagus and lung

K Suzuki, Y Daigo, S Fukuda, T Tokino, M Isomura, K Isono, B Wainwright, Y Nakamura

JAPANESE JOURNAL OF CANCER RESEARCH ( ELSEVIER SCI IRELAND LTD ) 88 ( 3 ) 225 - 228 1997.03

The high frequency of loss of heterozygosity that has been observed on the distal region of the long arm of chromosome 9 in squamous cell carcinomas of esophagus, lung, uterus, and head and neck indicates the presence of a tumor suppressor gene(s) in this region. To investigate the possible role of the PTC gene on chromosome 9q22.3, that was identified as the cause of nevoid basal cell carcinoma syndrome, during carcinogenesis in esophagus and lung, we examined 20 esophageal squamous cell carcinomas and 10 squamous cell carcinomas of the lung for mutations in any coding exon of PTC. Using single-strand conformation polymorphism and direct sequencing, we detected no mutations other than two non-deleterious polymorphisms. Our results suggest that inactivation of some tumor suppressor gene(s) on 9q other than PTC contributes to the development of squamous cell carcinomas in these tissues.

Mutational analysis of the hMLH1 gene using an automated two-dimensional DNA typing system

S Sasaki, T Tokino, T Miyatsu, T Muto, Y Nakamura

HUMAN MUTATION ( WILEY-LISS ) 9 ( 2 ) 164 - 171 1997

Searching for mutations in DNA mismatch repair genes is important not only for presymptomatic diagnosis, but also for documenting the spectrum of mutations among families carrying predispositions to hereditary nonpolyposis colorectal cancer (HNPCC). We utilized an automated two-dimensional DNA typing system for mutational analysis of the hMLH1 gene and established optimal conditions for application of the technique to analysis of hMLH1. This approach enabled us to visualize 21 spots covering all 19 coding exons on a single gel and to envisage whether and where: any mutations existed. All mutations that we had detected previously by other means in a panel of HNPCC patients and in one patient with sporadic endometrial cancer were also detectable by this method. Furthermore, using the 2-D system, we screened the entire coding regions of the hMLH1 gene in DNAs isolated from affected individuals belonging to two large HNPCC kindreds and four HNPCC-like kindreds, and from four patients with multiple primary cancers as well as eight sporadic colorectal cancers with replication error (RER)-positive phenotypes. We detected novel germline mutations in one HNPCC proband and one sporadic colorectal cancer with the RER-positive phenotype and one polymorphism in two HNPCC-like kindreds. This new diagnostic method is applicable to mutation of analysis of any disease causing gene, and it offers a major improvement over current approaches. (C) 1997 Wiley-Liss, Inc.

Cloning and characterization of BAI2 and BAI3, novel genes homologous to brain-specific angiogenesis inhibitor 1 (BAI1)

T Shiratsuchi, H Nishimori, H Ichise, Y Nakamura, T Tokino

CYTOGENETICS AND CELL GENETICS ( KARGER ) 79 ( 1-2 ) 103 - 108 1997

We have identified two novel human genes homologous to BAI1 (brain-specific angiogenesis inhibitor 1), an angiogenesis inhibitor that is a candidate for involvement in development of glioblastoma. Like BAI1, these two genes, designated BAI2 and BAI3, were specifically expressed in brain, and are likely to be expressed in the same type of cells. However, in spite of similar tissue specificity among the three BAI genes, only BAI1 is transcriptionally regulated by p53. BAI3 expression was absent in two of nine glioblastoma cell lines examined and was significantly reduced in three of the remaining seven. These data suggest that members of this novel gene family may play important roles in suppression of glioblastoma. BAI1, BAI2 and BAI3 were mapped to 8q24, 1p35 and 6q12, respectively.

Isolation of a novel GPI-anchored gene specifically regulated by p53; Correlation between its expression and anti-cancer drug sensitivity

T Furuhata, T Tokino, T Urano, Y Nakamura

ONCOGENE ( STOCKTON PRESS ) 13 ( 9 ) 1965 - 1970 1996.11

We have identified a novel gene inducible by wild-type p53. A significant correlation between expression of this gene and p53 status in cells derived from esophageal cancers indicated that this gene is likely to be specifically regulated in a p53-dependent manner. As the predicted amino acid sequence showed a high degree of homology to the family of glycosyl-phosphatidylinositol (GPI)-anchored membrane proteins, we termed this gene GML (GPI-anchored molecule-like protein). Introduction of GML cDNA suppressed the growth of esophageal cancer cells in culture. A correlation between the presence of GML expression and the sensitivity of esophageal cancer cells to anti-cancer drugs implied that the gene product plays a significant role in the apoptotic pathway or cell-cycle regulation induced by p53 after DNA damage.

Cooperation of the tumour suppressors IRF-1 and p53 in response to DNA damage

N Tanaka, M Ishihara, MS Lamphier, H Nozawa, T Matsuyama, TW Mak, S Aizawa, Takashi Tokino, M Oren, T Taniguchi

NATURE ( MACMILLAN MAGAZINES LTD ) 382 ( 6594 ) 816 - 818 1996.08

NORMALLY growing cells promptly cease DNA synthesis when exposed to genotoxic stresses, such as radiation, and this cell-cycle arrest prevents the accumulation of mutations(1,2). The transcription factor interferon regulatory factor (IRF)-1 is essential for the regulation of the interferon system(3-5), inhibits cell growth, and manifests tumour-suppressor activities(6,7). Here we show that mouse embryonic fibroblasts (EFs) lacking LRF-1 are deficient in their ability to undergo DNA-damage-induced cell-cycle arrest. A similar phenotype has been observed in EFs lacking the tumour suppressor p53 (refs 8, 9), although the expression of IRF-1 and p53 are independent of one another. Furthermore, we show that transcriptional induction of the gene encoding p21 (WAF1, CIP1)(10-12), a cell-cycle inhibitor, by gamma-irradiation is dependent on both p53 and IRF-1, and that the p21 promoter is activated, either directly or indirectly, by both in a transient cotransfection assay. These two tumour-suppressor transcription factors therefore converge functionally to regulate the cell cycle through the activation of a common target genes.

Characterization of the human p57(KIP2) gene: Alternative splicing, insertion deletion polymorphisms in VNTR sequences in the coding region, and mutational analysis

Takashi Tokino, T Urano, T Furuhata, M Matsushima, T Miyatsu, S Sasaki, Y Nakamura

HUMAN GENETICS ( SPRINGER VERLAG ) 97 ( 5 ) 625 - 631 1996.05

We have isolated human cDNA and genomic clones of a gene termed p57(KIP2), which is related to the p21(WAF1) and p27(KIP1) genes that encode inducible inhibitors of cyclin-dependent kinase activity. The p57 gene contains three GC-rich introns of 166 bp, 566 bp, and 83 bp, and two of the four exons correspond to coding regions. Alternative splicing generates the heterogeneity in the translational initiations. As this gene has been localized to chromosomal band 11p15.5, a region thought to be the location of a tumor suppressor gene(s) for carcinomas of the breast, bladder, and liver, we have examined a large number of tumors for genetic alterations of p57. Although no somatic mutation has been detected, we have found several normal variations in this gene, including four types of 12-bp in-frame deletions in the proline/alanine repeating domain, in which nearly 40 motifs, viz., 5'CCGGCC-3', are tandemly repeated.

Definition of the locus responsible for systemic carnitine deficiency within a 1.6-cM region of mouse chromosome 11 by detailed linkage analysis

K Okita, T Tokino, H Nishimori, K Miura, H Nikaido, J Hayakawa, A Ono, M Kuwajima, Y Matsuzawa, Y Nakamura

GENOMICS ( ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS ) 33 ( 2 ) 289 - 291 1996.04

Carnitine is an essential cofactor for oxidation of mitochondrial fatty acids. Carnitine deficiency results in failure of energy production by mitochondria and leads to metabolic encephalopathy, lipid-storage myopathy, and cardiomyopathy. The juvenile visceral steatosis (JVS) mouse, an animal model of systemic carnitine deficiency, inherits the JVS phenotype in autosomal recessive fashion, through a mutant allele mapped to mouse chromosome 11. As a step toward identifying the gene responsible for JVS by positional cloning, we attempted to refine the jvs locus in the mouse by detailed linkage analysis with 13 microsatellite markers, using 190 backcross progeny. Among the 13 loci tested, 5 (defined by markers D11Mit24, D11Mit111, D11Nds9, D11Mit86, and D11Mit23) showed no recombination, with a maximum lod score of 52.38, Our results implied that the jvs gene can be sought on mouse chromosome 11 within a genetic distance no greater than about 1.6 cM. (C) 1996 Academic Press, Inc.

Detailed deletion mapping in squamous cell carcinomas of the esophagus narrows a region containing a putative tumor suppressor gene to about 200 kilobases on distal chromosome 9q(1)

K Miura, K Suzuki, T Tokino, M Isomura, J Inazawa, S Matsuno, Y Nakamura

CANCER RESEARCH ( AMER ASSOC CANCER RESEARCH ) 56 ( 7 ) 1629 - 1634 1996.04

We previously reported definition of a region containing a putative tumor suppressor gene for esophageal squamous cell carcinoma within an similar to 4-cM genomic segment at 9q31-q32. We have investigated this region further using six new microsatellite markers isolated from yeast artificial chromosome clones covering the deleted region and have narrowly defined the commonly deleted region to a segment between two loci, KM9.1 and D9S177, On the basis of the contig map of cosmid and yeast artificial chromosome clones, we estimate the physical size of the region of interest to be about 200 kb, Because the distal 9q region also has been implicated as the site of a tumor suppressor gene(s) related to squamous cell carcinomas of other tissues, our map provides useful information for attempts to identify a common gene for carcinomas of this cell type.

MAPPING THE MOUSE GENE FOR PRIMARY CARNITINE DEFICIENCY

K OKITA, T TOKINO, H NISHIMORI, H NIKAIDO, J HAYAKAWA, A OHNO, M KUWAJIMA, Y MATSUZAWA, Y NAKAMURA

AMERICAN JOURNAL OF HUMAN GENETICS ( UNIV CHICAGO PRESS ) 57 ( 4 ) 1890 - 1890 1995.10

Research paper, summary (international conference)

TOPOLOGICAL CONTROL OF P21(WAF1/CIP1) EXPRESSION IN NORMAL AND NEOPLASTIC TISSUES

WS ELDEIRY, Takashi Tokino, T WALDMAN, JD OLINER, VE VELCULESCU, M BURRELL, DE HILL, E HEALY, JL REES, HAMILTON, SR, KW KINZLER, B VOGELSTEIN

CANCER RESEARCH ( AMER ASSOC CANCER RESEARCH ) 55 ( 13 ) 2910 - 2919 1995.07

The p53-regulated gene product p21(WAF1/CIP1) is the prototype of a family of small proteins that negatively regulate the cell cycle. To learn more about p21(WAF1/CIP1) regulation in vivo, monoclonal antibodies were developed for immunohistochemistry. These revealed that p21(WAF1/CIP1) expression followed radiation-induced DNA damage in human skin in a pattern consistent with its regulation by p53. A detailed comparison of the human, rat, and mouse p21(WAF1/CIP1) promoter sequences revealed that this induction was probably mediated by conserved p53-binding sites upstream of the transcription start site. In unirradiated tissues, p21(WAF1/CIP1) expression was apparently independent of p53 and was observed in a variety of cell types. Moreover, there was a striking compartmentalization of p21(WAF1/CIP1) expression throughout the gastrointestinal tract that correlated with proliferation rather than differentiation. As epithelial cells migrated up the crypts, the Ki67-expressing proliferating compartment near the crypt base ended abruptly, with the coincident appearance of a nonproliferating compartment expressing p21(WAF1/CIP1). In colored neoplasms, this distinct compartmentalization was largely abrogated. Cell cycle inhibitors are thus subject to precise topological control, and escape from this regulation may be a critical feature of neoplastic transformation.

P53-DEPENDENT AND INDEPENDENT EXPRESSION OF P21 DURING CELL-GROWTH, DIFFERENTIATION, AND DNA-DAMAGE

KF MACLEOD, N SHERRY, G HANNON, D BEACH, T TOKINO, K KINZLER, B VOGELSTEIN, T JACKS

GENES & DEVELOPMENT ( COLD SPRING HARBOR LAB PRESS ) 9 ( 8 ) 935 - 944 1995.04

Expression of p21 has been shown to be up-regulated by the p53 tumor suppressor gene in vitro in response to DNA-damaging agents. However, p21 expression can be regulated independently of p53, and here we show that expression of p21 in various tissues during development and in the adult mouse occurs in the absence of p53 function. However, most tissues tested did require p53 for p21 induction following exposure of the whole animal to gamma irradiation. These results show that normal tissue expression of p21 to high levels is not dependent on p53 and confirm that induction of p21 by DNA-damaging agents does require p53. p21 is expressed upon differentiation of p53-deficient murine erythroleukemia (MEL) cells, and the kinetics of induction of p21 in this system suggest that it may be involved in the growth arrest that precedes terminal differentiation. The gene is up-regulated in mouse fibroblasts in response to serum restimulation but the kinetics and levels of induction differ between wild-type and mutant cells. Expression of p21 message following serum restimulation is superinducible by cycloheximide in wild-type but not in p53 deficient cells. The increases in p21 mRNA are reflected in changes in p21 protein levels. p21 expression also appears to be regulated at the post-transcriptional level because moderate increases in mRNA expression, during differentiation of MEL tells and upon serum restimulation of fibroblasts, are followed by large increases in protein levels, Regulation of the mouse p21 promoter by p53 depends on two critical p53-binding sites located 1.95 and 2.85 kb upstream from the transcriptional initiation site. The sequences mediating serum responsiveness of the promoter map to a region containing the proximal p53 site. p53 appears to play a critical role in p21 induction following DNA damage. Moreover, p21 can be regulated independently of p53 in several situations including during normal tissue development, following serum stimulation, and during cellular differentiation.

50 NOVEL SEQUENCE-TAGGED SITES (STSS) ON HUMAN-CHROMOSOME 11Q13.4-]Q25 IDENTIFIED FROM MICROCLONES GENERATED BY MICRODISSECTION

H SOEJIMA, K YOSHIURA, T TAMURA, T TOKINO, Y NAKAMURA, N NIIKAWA, Y JINNO

CYTOGENETICS AND CELL GENETICS ( KARGER ) 70 ( 1-2 ) 108 - 111 1995

Fifty sequence-tagged sites (STSs) from human chromosome region, 11q13.4-->q25 were identified in microdissection-generated microclones. From a total of 124 single-copy microclones obtained from regions 11q14-->q22 and 11q23-->q25, 59 were sequenced, and a PCR primer pair was designed for each of them. Fifty of the 59 dones were mapped to regions ranging from 11q13.4 to 11q25 by means of PCR on a somatic hybrid cell panel with various deletions of the long arm of human chromosome 11. These STSs will contribute to the construction of physical cosmid/YAC maps representing the chromosome regions. They are also useful for analysis of chromosome aberrations, such as translocations, inversions or marker chromosomes.

P53 TAGGED SITES FROM HUMAN GENOMIC DNA

T TOKINO, S THIAGALINGAM, WS ELDEIRY, T WALDMAN, KW KINZLER, B VOGELSTEIN

HUMAN MOLECULAR GENETICS ( OXFORD UNIV PRESS UNITED KINGDOM ) 3 ( 9 ) 1537 - 1542 1994.09

The product of the tumor suppressor gene p53 binds to DNA and activates transcription from promoters containing its consensus binding site. This activity has been hypothesized to be responsible for its biological effects. However, the total number and nature of human genomic sites with which p53 can functionally interact is unknown. In this paper, we have used a Saccharomyces cerevisiae-based screen to identify human genomic sequences that activate transcription from an adjacent reporter gene in a p53-dependent manner (p53-tagged sites, PTS). Fifty-seven different PTS were identified, and the total number of such sites in the human genome was predicted to be between 200 and 300. Almost all contained two adjacent copies of the previously defined consensus 5'-PuPuPuC(A/T)(T/A)GPyPyPy-3'. Spacing between the copies was found to be critical for sequence-specific transcriptional activation in vivo. These results further refine the nature of the genomic sequences likely to be most important for p53-mediated tumor suppression.

SEQUENCE-SPECIFIC TRANSCRIPTIONAL ACTIVATION IS ESSENTIAL FOR GROWTH SUPPRESSION BY P53

JA PIETENPOL, T TOKINO, S THIAGALINGAM, WS ELDEIRY, KW KINZLER, B VOGELSTEIN

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA ( NATL ACAD SCIENCES ) 91 ( 6 ) 1998 - 2002 1994.03

Although several biochemical features of p53 have been described, their relationship to tumor suppression remains uncertain. We have compared the ability of p53-derived proteins to act as sequence-specific transcriptional (SST) activators with their ability to suppress tumor cell growth, using an improved growth-suppression assay. Both naturally occurring and in vitro derived mutations that abrogated the SST activity of p53 lost the ability to suppress tumor cell growth. Additionally, the N- and C-terminal ends of p53 were shown to be functionally replaceable with foreign transactivation and dimerization domains, respectively, with concordant preservation of both SST and tumor-suppressive properties. Only the central region of p53, conferring specific DNA binding, was required to suppress growth by such hybrid proteins. The SST activity of p53 thus appeared to be essential for the protein to function as a tumor suppressor.

HIGH-RESOLUTION ORDERING OF DNA MARKERS BY MULTICOLOR FLUORESCENT IN-SITU HYBRIDIZATION OF PROPHASE CHROMOSOMES

J INAZAWA, T ARIYAMA, T TOKINO, A TANIGAMI, Y NAKAMURA, T ABE

CYTOGENETICS AND CELL GENETICS ( KARGER ) 65 ( 1-2 ) 130 - 135 1994

To improve resolution for physical ordering of adjacent DNA loci, prophase chromosomes were used for multi-color fluorescent in situ hybridization (FISH). The prophase chromosomes were prepared from cultured lymphocytes by a thymidine synchronization, bromodeoxyuridine release technique and then treating the synchronized cultures with topoisomerase II inhibitors ICRF154 or ICRF193. Almost all mitotic figures exhibited highly elongated prophase chromosomes without significant reduction of the mitotic index. Using multi-color FISH with these prophase chromosomes, we were able to distinguish signals for loci separated by as little as 50 kb, and determine their orientation. Furthermore, using this prophase ordering system, we confirmed the linear order and defined the orientation of seven cosmid markers within a 360-kb region surrounding D10S102, a locus that is closely linked to the disease locus in families segregating an allele causing multiple endocrine neoplasia IIA (MEN2A). This prophase FISH system, by rapidly and precisely providing the linear order of loci that are very close, can expedite construction of fine cytogenetic maps and contribute to positional-cloning studies in which the precise ordering of DNA loci in a target region is critical.

WAF1, A POTENTIAL MEDIATOR OF P53 TUMOR SUPPRESSION

WS ELDEIRY, T TOKINO, VE VELCULESCU, DB LEVY, R PARSONS, JM TRENT, D LIN, WE MERCER, KW KINZLER, B VOGELSTEIN

CELL ( CELL PRESS ) 75 ( 4 ) 817 - 825 1993.11

The ability of p53 to activate transcription from specific sequences suggests that genes induced by p53 may mediate its biological role as a tumor suppressor. Using a subtractive hybridization approach, we identified a gene, named WAF1, whose induction was associated with wild-type but not mutant p53 gene expression in a human brain tumor cell line. The WAF1 gene was localized to chromosome 6p21.2, and its sequence, structure, and activation by p53 was conserved in rodents. Introduction of WAF1 cDNA suppressed the growth of human brain, lung, and colon tumor cells in culture. Using a yeast enhancer trap, a p53-binding site was identified 2.4 kb upstream of WAF1 coding sequences. The WAF1 promoter, including this p53-binding site, conferred p53-dependent inducibility upon a heterologous reporter gene. These studies define a gene whose expression is directly induced by p53 and that could be an important mediator of p53-dependent tumor growth suppression.

ABSENCE OF GERMLINE PROHIBITIN MUTATIONS IN EARLY-ONSET BREAST-CANCER

T TOKINO, N DAVIDSON, K HELZLSOUER, B ZEHNBAUER, Y NAKAMURA, B VOGELSTEIN, D SIDRANSKY

INTERNATIONAL JOURNAL OF ONCOLOGY ( INT JOURNAL ONCOLOGY ) 3 ( 4 ) 769 - 772 1993.10

The prohibitin gene, mapped to chromosome 17q21 is a candidate tumor suppressor gene. Because several somatic mutations of prohibitin were detected in breast tumors, we sought to identify potential carriers of prohibitin gene mutations in selected patients with early onset breast cancer. Seventy-six patients who developed breast cancer at or before the age of 40 were examined for mutations of prohibitin within the most conserved regions of the gene. Additionally, all exons of the prohibitin gene were examined in a subgroup of patients with a history of first-degree relatives with primary breast cancer. While several polymorphisms were noted, no inherited mutations of prohibitin were identified. This study suggests that prohibitin is not an early onset breast cancer susceptibility gene and additional studies will be necessary to understand its role in sporadic breast cancer progression.

IDENTIFICATION AND CHARACTERIZATION OF A 9-KB SOMATIC HOMOZYGOUS DELETION AT 3P21.3-P22 IN A LUNG-CANCER CELL-LINE

K YAMAKAWA, Y MURATA, M TAMARI, T TAKAHASHI, Y HORIO, K HIBI, S YOKOYAMA, J INAZAWA, T TAKAHASI, Y NAKAMURA

AMERICAN JOURNAL OF HUMAN GENETICS ( UNIV CHICAGO PRESS ) 53 ( 3 ) 390 - 390 1993.09

Research paper, summary (international conference)

P53 MUTATION AND MDM2 AMPLIFICATION IN HUMAN SOFT-TISSUE SARCOMAS

FS LEACH, T TOKINO, P MELTZER, M BURRELL, JD OLINER, S SMITH, DE HILL, D SIDRANSKY, KW KINZLER, B VOGELSTEIN

CANCER RESEARCH ( AMER ASSOC CANCER RESEARCH ) 53 ( 10 ) 2231 - 2234 1993.05

Article, review, commentary, editorial, etc. (scientific journal)

The p53 and MDM2 genes were analyzed in 24 human soft tissue sarcomas (11 malignant fibrous histiocytomas and 13 liposarcomas). Alterations of p53, consisting of point mutations, deletions, or overexpression, were detected in one-third (8 of 24) of the sarcomas. MDM2 gene amplification was detected in another 8 tumors, but no tumor contained an alteration of both genes. Monoclonal antibodies reactive with the human MDM2 gene product were developed, and immunohistochemical analysis revealed nuclear localization and overexpression of MDM2 in those tumors with amplified MDM2 genes. These data support the hypothesis that p53 and MDM2 genetic alterations are alternative mechanisms for inactivating the same regulatory pathway for suppressing cell growth.

MAPPING THE BREAKPOINT OF A CONSTITUTIONAL TRANSLOCATION ON CHROMOSOME-22 IN A PATIENT WITH NF2

E ARAI, T TOKINO, T IMAI, J INAZAWA, T IKEUCHI, A TONOMURA, Y NAKAMURA

GENES CHROMOSOMES & CANCER ( WILEY-LISS ) 6 ( 4 ) 235 - 238 1993.04

Neurofibromatosis type 2 (NF2) is an autosomal dominant disorder characterized by development of bilateral acoustic neurinomas and increased incidence of meningiomas. Frequent losses of 1 allele of chromosome 22 in neurinomas and meningiomas has indicated that the gene responsible for NF2 functions as a tumor suppressor. Although the NF2 gene has been mapped within a 13 cM region between D22S1 and D22S28 by linkage analysis, its location with respect to D22S15 is uncertain. We previously reported an NF2 patient with a constitutional balanced translocation t(4;22) (q12;q12.2); the NF2 gene is probably disrupted at the breakpoint. To define the location of this breakpoint on chromosome 22, we performed fluorescence in situ hybridization (FISH) with DNA markers in the NF2 region and determined the physical order of 5 loci: D22S1-NF2-LIF-D22S15-D22S32.

ASSIGNMENT OF 54 COSMID CLONES TO 5 REGIONS OF CHROMOSOME-10

SE MOLE, MS JACKSON, T TOKINO, Y NAKAMURA, BAJ PONDER

GENOMICS ( ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS ) 15 ( 2 ) 457 - 458 1993.02

Article, review, commentary, editorial, etc. (scientific journal)

IDENTIFICATION AND CHARACTERIZATION OF A CDNA, WHICH IS HIGHLY HOMOLOGOUS TO THE RIBONUCLEOPROTEIN GENE, FROM A LOCUS (D10S102) CLOSELY LINKED TO MEN2 (MULTIPLE ENDOCRINE NEOPLASIA TYPE-2)

S TAKIGUCHI, T TOKINO, T IMAI, A TANIGAMI, K KOYAMA, Y NAKAMURA

CYTOGENETICS AND CELL GENETICS ( KARGER ) 64 ( 2 ) 128 - 130 1993

We previously reported a nearly 1-Mb contig of cosmid and YAC clones at D10S102, a locus that is tightly linked to the gene responsible for multiple endocrine neoplasia type 2 (MEN2A). Subclones of 159 genomic EcoRI fragments in the contig were tested on ''zoo blot'' to detect DNA sequences conserved in various species. Nine fragments found to contain conserved sequences were used as probes for screening cDNA libraries. One of these probes, y300-pL50, recognized a cDNA from a human fetal brain library. The cDNA encoded a protein consisting of 270 amino acids, whose predicted sequence has a high degree of homology to heterogeneous nuclear ribonuclear protein. This expressed sequence may be a candidate for the gene responsible for MEN2A.

Mapping of human chromosome 3 and its application to human diseases

K. Yamakawa, Y. Nakamura, E. Takahashi

Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme 38 ( 3 ) 215 - 227 1993

Book review, literature introduction, etc.

IDENTIFICATION OF 57 CONVENTIONAL RFLP AND 6 VNTR SYSTEMS WITH 32 DNA CLONES ON CHROMOSOME-11P15

K TAKITA, A TANIGAMI, T TOKINO, C JONES, Y NAKAMURA

GENOMICS ( ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS ) 13 ( 4 ) 1296 - 1299 1992.08

A HIGH-RESOLUTION CYTOGENETIC MAP OF 168-COSMID DNA MARKERS FOR HUMAN CHROMOSOME-11

TA HORI, E TAKAHASHI, A TANIGAMI, T TOKINO, Y NAKAMURA

GENOMICS ( ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS ) 13 ( 1 ) 129 - 133 1992.05

INHERITED P53 GENE-MUTATIONS IN BREAST-CANCER

D SIDRANSKY, T TOKINO, K HELZLSOUER, B ZEHNBAUER, G RAUSCH, B SHELTON, L PRESTIGIACOMO, B VOGELSTEIN, N DAVIDSON

CANCER RESEARCH ( AMER ASSOC CANCER RESEARCH ) 52 ( 10 ) 2984 - 2986 1992.05

Article, review, commentary, editorial, etc. (scientific journal)

Recent evidence has implicated germ-line mutations of the p53 gene as the cause of cancer susceptibility in the Li-Fraumeni syndrome, associated with the development of breast cancer and other neoplasms. Furthermore, somatic mutations of the p53 gene have been detected in a high percentage of non-familial breast cancers. We therefore sought to identify potential carriers of p53 gene mutations in a cohort of patients with early onset breast cancer. We examined 126 consecutive patients who developed breast cancer at or before the age of 40 for mutations of p53 within conserved regions of the gene. One patient with an inherited germ-line mutation of the p53 gene was identified but the functional significance of this mutation was not clear. It thus appears that only a small percentage of patients with breast cancer under the age of 40 carry germ-line mutations of the p53 gene, an observation which has implications for potential screening and risk assessment in such patients.

DETAILED ANALYSIS OF AN AMPLIFIED REGION AT CHROMOSOME-11Q13 IN MALIGNANT-TUMORS

A TANIGAMI, Takashi Tokino, K TAKITA, M UEDA, F KASUMI, Y NAKAMURA

GENOMICS ( ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS ) 13 ( 1 ) 21 - 24 1992.05

IDENTIFICATION OF RAS ONCOGENE MUTATIONS IN THE STOOL OF PATIENTS WITH CURABLE COLORECTAL TUMORS

D SIDRANSKY, Takashi Tokino, HAMILTON, SR, KW KINZLER, B LEVIN, P FROST, B VOGELSTEIN

SCIENCE ( AMER ASSOC ADVANCEMENT SCIENCE ) 256 ( 5053 ) 102 - 105 1992.04

Colorectal (CR) tumors are usually curable if detected before metastasis. Because genetic alterations are associated with the development of these tumors, mutant genes may be found in the stool of individuals with CR neoplasms. The stools of nine patients whose tumors contained mutations of K-ras were analyzed. In eight of the nine cases, the ras mutations were detectable in DNA purified from the stool. These patients included those with benign and malignant neoplasms from proximal and distal colonic epithelium. Thus, colorectal tumors can be detected by a noninvasive method based on the molecular pathogenesis of the disease.

ASSOCIATION BETWEEN CHROMOSOME-11Q13 AMPLIFICATION AND PROGNOSIS OF PATIENTS WITH ESOPHAGEAL CARCINOMAS

M MORI, Takashi Tokino, A YANAGISAWA, M KANAMORI, Y KATO, Y NAKAMURA

EUROPEAN JOURNAL OF CANCER ( PERGAMON-ELSEVIER SCIENCE LTD ) 28A ( 4-5 ) 755 - 757 1992.04

Ninety pairs of normal and tumour tissue DNAs were isolated from paraffin-embedded blocks of advanced oesophageal carcinoma cases and examined for gene amplification at chromosome 11q13 by dot-blot hybridisation using the int-2 gene as a probe. 22 of 90 carcinomas (24%) showed more than two times amplification. Although no significant correlation was observed between gene amplification and histological type or metastasis to lymph node, a tendency for deeper invasion to be associated with more frequent amplification was observed. In relation to prognosis, patients with amplification had a lower survival rate than those without amplification. This tendency was evident both in the group with well differentiated type carcinoma and in the group which had no metastasis to lymph node. Thus, gene amplification of the int-2 locus may be a useful prognostic factor.

PREDICTIVE TESTING FOR MULTIPLE ENDOCRINE NEOPLASIA TYPE-1 USING DNA POLYMORPHISMS

C LARSSON, J SHEPHERD, Y NAKAMURA, C BLOMBERG, G WEBER, B WERELIUS, N HAYWARD, B TEH, T TOKINO, B SEIZINGER, B SKOGSEID, K OBERG, M NORDENSKJOLD

JOURNAL OF CLINICAL INVESTIGATION ( ROCKEFELLER UNIV PRESS ) 89 ( 4 ) 1344 - 1349 1992.04

Mulitple endocrine neoplasia type 1 (MEN1) is an autosomal dominantly inherited predisposition to neoplastic lesions of the parathyroids, pancreas, and the pituitary. We have previously located the predisposing genetic defect to the long arm of chromosome 11 by genetic linkage. In this study, 124 members of six MEN1 families, including 59 affected individuals, were genotyped for restriction fragment length polymorphisms with different DNA probes, and the genetic linkage between these marker systems and MEN1 was determined. 13 marker systems (17 DNA probes) were found to be linked to MEN1. These markers are located within a region on chromosome 11 spanning 14% meiotic recombinations, with the MEN1 locus in the middle. Four of the marker systems are on the centromeric side of MEN1, and four on the telomeric side, based on meiotic crossovers. The remaining five DNA probes are closely linked to MEN1, with no crossovers in our set of families. The 13 marker systems can be used for an accurate and reliable premorbid test for MEN1. In most clinical situations it is possible to identify a haplotype of this part of chromosome 11 with the mutant MEN1 allele in the middle. The calculated predictive accuracy is > 99.5% if three such marker systems are informative. Therefore, genetic linkage testing can be used for informed genetic counseling in MEN1 families, and to avoid unnecessary biochemical screening programs.

THE 11Q23 BREAKPOINT IN ACUTE-LEUKEMIA WITH T(11-19)(Q23-P13) IS DISTAL TO THOSE OF T(4-11), T(6-11) AND T(9-11)

D CHERIF, H DERSARKISSIAN, J DERRE, T TOKINO, Y NAKAMURA, R BERGER

GENES CHROMOSOMES & CANCER ( WILEY-LISS ) 4 ( 2 ) 107 - 112 1992.03

Thirteen cosmid probes were mapped on the long arm of chromosome 11 between 11q22 and 11q24 by nonradioactive in situ hybridization. Starting with these localizations and those of other probes mapped to 11q23, four acute leukemias with translocations involving 11q23 were studied with the same method. The translocation breakpoints of the t(4;11)(q21;q23), t(6;11)(q27;q23), t(9;11)(p21-p22;q23), and t(11;19)(q23;p13) were confirmed to be distal to CD3D. The probe cC111-304 was proximal to the t(11;19) breakpoint while distal to the breakpoints of the other rearrangements. In view of the diversity of chromosomal abnormalities involving band 11q23, our finding extends the molecular heterogeneity of the breakpoint localization in leukemias with rearrangements involving 11q23.

31 NEW RFLP SYSTEMS DETECTED BY 24 DNA MARKERS ON HUMAN CHROMOSOME-10

T TOKINO, S TAKIGUCHI, A TANIGAMI, T BRAGG, C JONES, Y NAKAMURA

GENOMICS ( ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS ) 12 ( 2 ) 401 - 402 1992.02

Article, review, commentary, editorial, etc. (scientific journal)

PHYSICAL MAPPING OF A 950-KB REGION SURROUNDING A LOCUS (D10S102) TIGHTLY LINKED TO THE MEN2A GENE

T TOKINO, T IMAI, A TANIGAMI, S TAKIGUCHI, Y NAKAMURA

GENOMICS ( ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS ) 12 ( 2 ) 394 - 400 1992.02

MAPPING OF 262 DNA MARKERS INTO 24 INTERVALS ON HUMAN CHROMOSOME-11

A TANIGAMI, Takashi Tokino, S TAKIGUCHI, M MORI, T GLASER, JW PARK, C JONES, Y NAKAMURA

AMERICAN JOURNAL OF HUMAN GENETICS ( UNIV CHICAGO PRESS ) 50 ( 1 ) 56 - 64 1992.01

We have extended our mapping effort on human chromosome 11 to encompass a total of 262 DNA markers, which have been mapped into 24 intervals on chromosome 11; 123 of the markers reveal RFLPs. These clones are scattered throughout the chromosome, although some clustering occurs in R-positive bands (p15.1, p11.2, q13, and q23.3). Fifty-two of the markers were found to contain DNA sequences conserved in Chinese hamster, and some of these 52 also cross-hybridized with DNA from other mammals and/or chicken. As the length of chromosome 11 is estimated at nearly 130 cM, the average distance between RFLP markers is roughly 1 cM. The large panel of DNA markers on our map should contribute to investigations of hereditary diseases on this chromosome, and it will also provide reagents for constructing either fine-scale linkage and physical maps or contig maps of cosmids or yeast artificial chromosomes.

ISOLATION AND MAPPING OF 68 RFLP MARKERS ON HUMAN CHROMOSOME-6

S SAITO, K OKUI, Takashi Tokino, M OSHIMURA, Y NAKAMURA

AMERICAN JOURNAL OF HUMAN GENETICS ( UNIV CHICAGO PRESS ) 50 ( 1 ) 65 - 70 1992.01

We have isolated 68 new RFLP markers on human chromosome 6. Of these, 64 were localized on chromosomal bands by the fluorescent in-situ hybridization (FISH) method, 25 on the short arm and 39 on the long arm. Their distribution was uneven; the markers were localized predominantly in regions of R-positive banding. Eleven markers defined VNTR loci. This expanded collection of DNA markers will contribute to high-resolution linkage mapping of genes causing inherited disorders and will provide useful reagents for isolation of putative tumor-suppressor genes on chromosome 6 that appear to be involved in malignancies. Furthermore, the new markers will be guideposts for detailed linkage and physical maps of this chromosome.

CHROMOSOMAL SITES FOR HEPATITIS-B VIRUS INTEGRATION IN HUMAN HEPATOCELLULAR-CARCINOMA

T TOKINO, K MATSUBARA

JOURNAL OF VIROLOGY ( AMER SOC MICROBIOLOGY ) 65 ( 12 ) 6761 - 6764 1991.12

The discovery that hepatitis B'virus (HBV) integrates into host chromosomes raises the question of whether such viral DNA integration correlates directly with the activation of specific oncogenes or the inactivation of anti-oncogenes. To obtain insight into this problem, we randomly collected HBV integrant samples from different human hepatocellular carcinomas and identified the site of chromosomal integration by using in situ hybridization and/or linkage analysis with the flanking cellular DNAs as probes. Our findings did not specifically identify particular HBV DNA integration sites in chromosomes, although chromosomes 11 and 17 seemed to have more than the average number of integrants.

CHROMOSOME DELETIONS ASSOCIATED WITH HEPATITIS-B VIRUS INTEGRATION

T TOKINO, H TAMURA, N HORI, K MATSUBARA

VIROLOGY ( ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS ) 185 ( 2 ) 879 - 882 1991.12

Article, review, commentary, editorial, etc. (scientific journal)

CLINICAL EPIDEMIOLOGY AND EVALUATION OF GENE AMPLIFICATION AT CHROMOSOME LLQ13 IN THE 90 CASES OF ESOPHAGEAL CARCINOMAS

M KANAMORI, M MORI, T TOKINO, A YANAGISAWA, Y KATO, Y NAKAMURA

AMERICAN JOURNAL OF HUMAN GENETICS ( UNIV CHICAGO PRESS ) 49 ( 4 ) 121 - 121 1991.10

Research paper, summary (international conference)

ISOLATION AND MAPPING OF 62 NEW RFLP MARKERS ON HUMAN CHROMOSOME-11

T TOKINO, E TAKAHASHI, M MORI, A TANIGAMI, T GLASER, JW PARK, C JONES, TA HORI, Y NAKAMURA

AMERICAN JOURNAL OF HUMAN GENETICS ( UNIV CHICAGO PRESS ) 48 ( 2 ) 258 - 268 1991.02

To obtain new RFLP markers on human chromosome 11 for a high-resolution map, we constructed a cosmid library from a Chinese hamster X human somatic hybrid cell line that retains only human chromosome 11 in a Chinese hamster genomic background. A total of 3,500 cosmids were isolated by colony hybridization with labeled human genomic DNA. DNA was prepared from 130 of these cosmid clones and examined for RFLP. In 62 of them, polymorphism was detected with one or more enzymes; four RFLPs were VNTR systems. All polymorphic clones were assigned to one of 22 intervals obtained by mapping on a deletion panel of 15 somatic hybrid cell lines containing parts of chromosome 11; 11 clones were finely mapped by in situ hybridization. Although RFLP markers were scattered on the whole chromosome, they were found predominantly in the regions of R-banding. These DNA markers will contribute to fine mapping of genes causing inherited disorders and tumor-suppressor genes that reside on chromosome 11. Furthermore, as one-third of the cosmid clones revealed a band or bands in Chinese hamster DNA, indicating sequence conservation, this subset of clones may be useful for isolating biologically important genes on chromosome 11.

ALLELOTYPE STUDY OF PRIMARY HEPATOCELLULAR-CARCINOMA

M FUJIMORI, T TOKINO, O HINO, T KITAGAWA, T IMAMURA, E OKAMOTO, M MITSUNOBU, T ISHIKAWA, H NAKAGAMA, H HARADA, M YAGURA, K MATSUBARA, Y NAKAMURA

CANCER RESEARCH ( AMER ASSOC CANCER RESEARCH ) 51 ( 1 ) 89 - 93 1991.01

Accumulation of mutations in oncogenes and tumor suppressor genes transforms a normal cell to a malignant cell by allowing it to escape from normal control of growth. In order to learn (a) how many tumor suppressor genes are involved in the tumor progression of hepatocellular carcinoma (b) whether there is any association among allelic losses of chromosomes, or (c) whether integration of hepatitis B virus into host DNA influences any particular chromosomal losses, we have examined loss of heterozygosity with 44 restriction fragment length polymorphism markers in 46 cases of hepatocellular carcinoma. The markers represented all chromosomal arms except 5p, 8p, 9p, 18p, and acrocentric chromosomes. Allelic losses in tumors indicated that five tumor suppressor genes, located on chromosomes 5q, 10q, 11p, 16q, and 17p, may be involved in this cancer. However, no significant associations were observed among the various allelic losses or between the integration of hepatitis B virus and chromosomal losses. Furthermore, a deletion map for chromosome 16q indicated the localization of a tumor suppressor gene between q22 and q24 and that for chromosome 17p suggested the existence of a second tumor suppressor gene in addition to the p53 gene.

TOWARDS A COMBINED MEIOTIC-BASED, RADIATION-HYBRID-BASED AND YAC-BASED PHYSICAL MAP OF CHROMOSOME-11Q22-Q23

MR JAMES, C JULIER, F CORNELIS, D CHERIF, R BERGER, Takashi Tokino, Y NAKAMURA, R MYERS, D COX, GM LATHROP, CW RICHARD

CYTOGENETICS AND CELL GENETICS ( KARGER ) 58 ( 3-4 ) 1964 - 1964 1991

Research paper, summary (international conference)

ISOLATION AND MAPPING OF 226 DNA MARKERS ON HUMAN CHROMOSOME-11

T TOKINO, A TANIGAMI, S TAKIGUCHI, T GLASER, JW PARK, C JONES, Y NAKAMURA

CYTOGENETICS AND CELL GENETICS ( KARGER ) 58 ( 3-4 ) 1972 - 1972 1991

Research paper, summary (international conference)

DEVELOPMENT OF POLYMORPHIC DINUCLEOTIDE REPEATS ON HUMAN CHROMOSOME-11

C DUBAY, J HAZAN, Takashi Tokino, Y NAKAMURA, CW RICHARD, D COX, MR JAMES, D CHERIF, M LATHROP, J WEISSENBACH

CYTOGENETICS AND CELL GENETICS ( KARGER ) 58 ( 3-4 ) 1958 - 1958 1991

Research paper, summary (international conference)

AMPLICON AT CHROMOSOME-11Q13 IN ESOPHAGEAL, BREAST AND HEPATOCELLULAR CARCINOMAS

A TANIGAMI, KI TAKITA, T TOKINO, Y NAKAMURA

CYTOGENETICS AND CELL GENETICS ( KARGER ) 58 ( 3-4 ) 1971 - 1971 1991

Research paper, summary (international conference)

A HIGH-RESOLUTION CYTOGENETIC MAP OF 149 MARKERS FOR HUMAN CHROMOSOME-11

TA HORI, EI TAKAHASHI, T TOKINO, Y NAKAMURA

CYTOGENETICS AND CELL GENETICS ( KARGER ) 58 ( 3-4 ) 1963 - 1963 1991

Research paper, summary (international conference)

PRECISE LOCALIZATION OF A GENE RESPONSIBLE FOR ATAXIATELANGIECTASIA ON CHROMOSOME-11Q

F CORNELIS, D CHERIF, M JAMES, T TOKINO, J DAVIES, D GIRAULT, C BERNARD, MF CROQUETTE, D THEAU, H AVETLOISEAU, J DERRE, M LITT, R BERGER, Y NAKAMURA, M LATHROP, C JULIER

CYTOGENETICS AND CELL GENETICS ( KARGER ) 58 ( 3-4 ) 1957 - 1957 1991

Research paper, summary (international conference)

A COMPARISON OF THE MOLECULAR-STRUCTURE OF INTEGRATED HEPATITIS-B VIRUS GENOMES IN HEPATOCELLULAR-CARCINOMA CELLS AND HEPATOCYTES DERIVED FROM THE SAME PATIENT

N OGATA, T TOKINO, T KAMIMURA, H ASAKURA

HEPATOLOGY ( W B SAUNDERS CO ) 11 ( 6 ) 1017 - 1023 1990.06

INTEGRATION OF HEPATITIS-B VIRUS-DNA AND ITS IMPLICATIONS FOR HEPATOCARCINOGENESIS

K MATSUBARA, T TOKINO

MOLECULAR BIOLOGY & MEDICINE ( ACADEMIC PRESS LTD ) 7 ( 3 ) 243 - 260 1990.06

CHROMOSOME REGIONAL MAPPING FOR THE HUMAN THYROID STIMULATING HORMONE BETA-SUBUNIT (TSHB) GENE

Takashi Tokino, Y HAYASHIZAKI, KI TAKATA, MC YOSHIDA, K MATSUBARA

CYTOGENETICS AND CELL GENETICS ( KARGER ) 53 ( 2-3 ) 140 - 143 1990

CO-AMPLIFICATION OF INTEGRATED HEPATITIS-B VIRUS-DNA AND TRANSFORMING GENE HST-1 IN A HEPATOCELLULAR-CARCINOMA

HATADA, I, T TOKINO, T OCHIYA, K MATSUBARA

ONCOGENE ( STOCKTON PRESS ) 3 ( 5 ) 537 - 540 1988.11

MICRODELETION ASSOCIATED WITH THE INTEGRATION PROCESS OF HEPATITIS-B VIRUS-DNA

T NAKAMURA, T TOKINO, T NAGAYA, K MATSUBARA

NUCLEIC ACIDS RESEARCH ( OXFORD UNIV PRESS UNITED KINGDOM ) 16 ( 11 ) 4865 - 4873 1988.06

REGIONAL LOCALIZATION OF THE LCA ONCOGENE TO HUMAN-CHROMOSOME REGION 2Q14-]Q21

T TOKINO, H SATOH, MC YOSHIDA, T OCHIYA, K MATSUBARA

CYTOGENETICS AND CELL GENETICS ( KARGER ) 48 ( 1 ) 63 - 64 1988

CHROMOSOMAL TRANSLOCATION AND INVERTED DUPLICATION ASSOCIATED WITH INTEGRATED HEPATITIS-B VIRUS IN HEPATOCELLULAR CARCINOMAS

T TOKINO, S FUKUSHIGE, T NAKAMURA, T NAGAYA, T MUROTSU, K SHIGA, N AOKI, K MATSUBARA

JOURNAL OF VIROLOGY ( AMER SOC MICROBIOLOGY ) 61 ( 12 ) 3848 - 3854 1987.12

THE MODE OF HEPATITIS-B VIRUS-DNA INTEGRATION IN CHROMOSOMES OF HUMAN HEPATOCELLULAR-CARCINOMA

T NAGAYA, T NAKAMURA, T TOKINO, T TSURIMOTO, M IMAI, T MAYUMI, K KAMINO, K YAMAMURA, K MATSUBARA

GENES & DEVELOPMENT ( COLD SPRING HARBOR LAB PRESS ) 1 ( 8 ) 773 - 782 1987.10

REPLICATION OF MINI-F PLASMID INVITRO PROMOTED BY PURIFIED E-PROTEIN

K MURAISO, T TOKINO, T MUROTSU, K MATSUBARA

MOLECULAR & GENERAL GENETICS ( SPRINGER VERLAG ) 206 ( 3 ) 519 - 521 1987.03

Article, review, commentary, editorial, etc. (scientific journal)

REPLICATION AND INTEGRATION OF HEPATITIS-B VIRUS GENOME

K MATSUBARA, T TSURIMOTO, K UEDA, O CHISAKA, T TOKINO

JOURNAL OF CELLULAR BIOCHEMISTRY ( WILEY-LISS ) 7 - 7 1987

Research paper, summary (international conference)

PURIFICATION AND PROPERTIES OF THE MINI-F PLASMID-ENCODED E-PROTEIN NEEDED FOR AUTONOMOUS REPLICATION CONTROL OF THE PLASMID

Takashi Tokino, T MUROTSU, K MATSUBARA

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA ( NATL ACAD SCIENCES ) 83 ( 12 ) 4109 - 4113 1986.06