論文 - 時野　隆至

次世代シークエンサーによる日本人 口腔扁平上皮癌におけるがん関連409遺伝子の変異解析

中垣 貴文, 佐々木 泰史, 井戸川 雅史, 小山 良太, 田村 みゆき, 福島 久代, 荻 和弘, 平塚 博義, 時野 隆至

日本癌学会総会記事 ( (一社)日本癌学会 )  76回   J - 1063  2017年09月

口腔扁平上皮癌の全エクソン解析

佐々木 泰史, 田村 みゆき, 中垣 貴文, 小山 良太, 井戸川 雅史, 荻 和弘, 平塚 博義, 仲瀬 裕志, 時野 隆至

日本癌学会総会記事 ( (一社)日本癌学会 )  76回   P - 3044  2017年09月

口腔扁平上皮癌に関与する長鎖非コードRNAの同定

西山 廣陽, 丸山 玲緒, 新沼 猛, 北嶋 洋志, 荻 和弘, 出張 裕也, 宮崎 晃亘, 甲斐 正広, 佐々木 泰史, 時野 隆至, 平塚 博義, 鈴木 拓

日本癌学会総会記事 ( (一社)日本癌学会 )  76回   P - 1292  2017年09月

ヒストンメチル化酵素DOT1Lは多発性骨髄腫の治療標的となりうる

石黒 一也, 佐々木 基, 若杉 英樹, 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

日本癌学会総会記事 ( (一社)日本癌学会 )  76回   P - 2244  2017年09月

Profiling cancer-related gene mutations in oral squamous cell carcinoma from Japanese patients by targeted amplicon sequencing.

Nakagaki T, Tamura M, Kobashi K, Koyama R, Fukushima H, Ohashi T, Idogawa M, Ogi K, Hiratsuka H, Tokino T, Sasaki Y

Oncotarget   8 ( 35 ) 59113 - 59122  2017年08月  [査読有り]  [国際誌]

　概要を見る

Somatic mutation analysis is a standard practice in the study of human cancers to identify mutations that cause therapeutic sensitization and resistance. We performed comprehensive genomic analyses that used PCR target enrichment and next-generation sequencing on Ion Proton semiconductor sequencers. Forty-seven oral squamous cell carcinoma (OSCC) samples and their corresponding noncancerous tissues were used for multiplex PCR amplification to obtain targeted coverage of the entire coding regions of 409 cancer-related genes (covered regions: 95.4% of total, 1.69 megabases of target sequence). The number of somatic mutations in 47 patients with OSCC ranged from 1 to 20 with a mean of 7.60. The most frequent mutations were in TP53 (61.7%), NOTCH1 (25.5%), CDKN2A (19.1%), SYNE1 (14.9%), PIK3CA (10.6%), ROS1 (10.6%), and TAF1L (10.6%). We also detected copy number variations (CNVs) in the segments of the genome that could be duplicated or deleted from deep sequencing data. Pathway assessment showed that the somatic aberrations within OSCC genomes are mainly involved in several important pathways, including cell cycle regulation and RTK-MAPK-PI3K. This study may enable better selection of therapies and deliver improved outcomes for OSCC patients when combined with clinical diagnostics.

DOI PubMed

Fledglings in p53 signaling network.

Tokino T, Idogawa M, Sasaki Y

Oncotarget   8 ( 34 ) 55768 - 55769  2017年08月  [査読有り]  [国際誌]

DOI PubMed

Mutation detection by target sequence analyses using tissue-specific panels in esophageal squamous cell carcinoma

Iwaya Takeshi, Endo Fumitaka, Kume Kohei, Sasaki Yasushi, Tokino Takashi, Nishizuka Satoshi

CANCER RESEARCH   77  2017年07月  [査読有り]

DOI

Long non-coding RNA NEAT1 is a transcriptional target of p53 and modulates p53-induced transactivation and tumor-suppressor function.

Masashi Idogawa, Tomoko Ohashi, Yasushi Sasaki, Hiroshi Nakase, Takashi Tokino

International journal of cancer   140 ( 12 ) 2785 - 2791  2017年06月  [査読有り]  [国際誌]

　概要を見る

p53 is one of the most important tumor suppressor genes, and the direct transcriptional targets of p53 must be explored to elucidate its functional mechanisms. Thus far, the p53 targets that have been primarily studied are protein-coding genes. Our previous study revealed that several long non-coding RNAs (lncRNAs) are direct transcriptional targets of p53, and knockdown of specific lncRNAs modulates p53-induced apoptosis. In this study, analysis of next-generation chromatin immunoprecipitation-sequencing (ChIP-seq) data for p53 revealed that the lncRNA NEAT1 is a direct transcriptional target of p53. The suppression of NEAT1 induction by p53 attenuates the inhibitory effect of p53 on cancer cell growth and also modulates gene transactivation, including that of many lncRNAs. Furthermore, low expression of NEAT1 is related to poor prognosis in several cancers. These results indicate that the induction of NEAT1 expression contributes to the tumor-suppressor function of p53 and suggest that p53 and NEAT1 constitute a transcriptional network contributing to various biological functions and tumor suppression.

DOI PubMed

p53 mediates the suppression of cancer cell invasion by inducing LIMA1/EPLIN.

Ohashi T, Idogawa M, Sasaki Y, Tokino T

Cancer letters   390   58 - 66  2017年04月  [査読有り]  [国際誌]

　概要を見る

The tumor suppressor gene p53 is frequently mutated in human cancer. p53 executes various functions, such as apoptosis induction and cell cycle arrest, by modulating transcriptional regulation. In this study, LIM domain and Actin-binding protein 1 (LIMA1) was identified as a target of the p53 family using a cDNA microarray. We also evaluated genome-wide occupancy of the p53 protein by performing chromatin immunoprecipitation-sequencing (ChIP-seq) and identified two p53 response elements in the LIMA1 gene. LIMA1 protein levels were increased by treatment with nutlin-3a, a small molecule that activates endogenous p53. In addition, LIMA1 expression was significantly downregulated in cancers compared with normal tissues. Knockdown of LIMA1 significantly enhanced cancer cell invasion and partially inhibited p53-induced suppression of cell invasion. Furthermore, low expression of LIMA1 in cancer patients correlated with decreased survival and poor prognosis. Thus, p53-induced LIMA1 inhibits cell invasion, and the downregulation of LIMA1 caused by p53 mutation results in decreased survival in cancer patients. Collectively, this study reveals the molecular mechanism of LIMA1 downregulation in various cancers and suggests that LIMA1 may be a novel prognostic predictor and a therapeutic target for cancer.

DOI PubMed

The effect of forced expression of mutated K-RAS gene on gastrointestinal cancer cell lines and the IGF-1R targeting therapy.

Matsunaga Y, Adachi Y, Sasaki Y, Koide H, Motoya M, Nosho K, Takagi H, Yamamoto H, Sasaki S, Arimura Y, Tokino T, Carbone DP, Imai K, Shinomura Y

Molecular carcinogenesis   56 ( 2 ) 515 - 526  2017年02月  [査読有り]  [国際誌]

　概要を見る

Mutation in K-RAS (K-RAS-MT) plays important roles in both cancer progression and resistance to anti-epidermal growth factor receptor (EGFR) therapy in gastrointestinal tumors. Insulin-like growth factor-1 receptor (IGF-1R) signaling is required for carcinogenicity and progression of many tumors as well. We have previously shown successful therapy for gastrointestinal cancer cell lines bearing a K-RAS mutation using an anti-IGF-1R monoclonal antibody. In this study, we sought to evaluate effects of forced K-RAS-MT expression on gastrointestinal cancer cell lines representing a possible second resistance mechanism for anti-EGFR therapy and IGF-1R-targeted therapy for these transfectants. We made stable transfectants of K-RAS-MT in two gastrointestinal cancer cell lines, colorectal RKO and pancreatic BxPC-3. We assessed the effect of forced expression of K-RAS-MT on proliferation, apoptosis, migration, and invasion in gastrointestinal cancer cells. Then we assessed anti-tumor effects of dominant negative IGF-1R (IGF-1R/dn) and an IGF-1R inhibitor, picropodophyllin, on the K-RAS-MT transfectants. Overexpression of K-RAS-MT in gastrointestinal cancer cell lines led to more aggressive phenotypes, with increased proliferation, decreased apoptosis, and increased motility and invasion. IGF-1R blockade suppressed cell growth, colony formation, migration, and invasion, and up-regulated chemotherapy-induced apoptosis of gastrointestinal cancer cells, even when K-RAS-MT was over-expressed. IGF-1R blockade inhibited the Akt pathway more than the extracellular signal-regulated kinase (ERK) pathway in the K-RAS-MT transfectants. IGF-1R/dn, moreover, inhibited the growth of murine xenografts expressing K-RAS-MT. Thus, K-RAS-MT might be important for progressive phonotype observed in gastrointestinal cancers. IGF-1R decoy is a candidate molecular therapeutic approach for gastrointestinal cancers even if K-RAS is mutated. © 2016 Wiley Periodicals, Inc.

DOI PubMed

Assessment of the quality of DNA from various formalin-fixed paraffin-embedded (FFPE) tissues and the use of this DNA for next-generation sequencing (NGS) with no artifactual mutation.

Einaga N, Yoshida A, Noda H, Suemitsu M, Nakayama Y, Sakurada A, Kawaji Y, Yamaguchi H, Sasaki Y, Tokino T, Esumi M

PloS one   12 ( 5 ) e0176280  2017年  [査読有り]  [国際誌]

　概要を見る

Formalin-fixed, paraffin-embedded (FFPE) tissues used for pathological diagnosis are valuable for studying cancer genomics. In particular, laser-capture microdissection of target cells determined by histopathology combined with FFPE tissue section immunohistochemistry (IHC) enables precise analysis by next-generation sequencing (NGS) of the genetic events occurring in cancer. The result is a new strategy for a pathological tool for cancer diagnosis: 'microgenomics'. To more conveniently and precisely perform microgenomics, we revealed by systematic analysis the following three details regarding FFPE DNA compared with paired frozen tissue DNA. 1) The best quality of FFPE DNA is obtained by tissue fixation with 10% neutral buffered formalin for 1 day and heat treatment of tissue lysates at 95°C for 30 minutes. 2) IHC staining of FFPE tissues decreases the quantity and quality of FFPE DNA to one-fourth, and antigen retrieval (at 120°C for 15 minutes, pH 6.0) is the major reason for this decrease. 3) FFPE DNA prepared as described herein is sufficient for NGS. For non-mutated tissue specimens, no artifactual mutation occurs during FFPE preparation, as shown by precise comparison of NGS of FFPE DNA and paired frozen tissue DNA followed by validation. These results demonstrate that even FFPE tissues used for routine clinical diagnosis can be utilized to obtain reliable NGS data if appropriate conditions of fixation and validation are applied.

DOI PubMed

p53ファミリー遺伝子の標的としてのBRMS1L(breast cancer metastasis suppressor 1 like、BRMS1-like)の同定と機能解析

小山 良太, 佐々木 泰史, 田村 みゆき, 中垣 貴文, 大箸 智子, 井戸川 雅史, 時野 隆至

日本癌学会総会記事 ( (一社)日本癌学会 )  75回   P - 2059  2016年10月

新規p53標的遺伝子LIMA1は癌細胞浸潤を抑制し低発現が予後不良と相関する

大箸 智子, 井戸川 雅史, 田村 みゆき, 中垣 貴文, 小山 良太, 佐々木 泰史, 時野 隆至

日本癌学会総会記事 ( (一社)日本癌学会 )  75回   J - 3027  2016年10月

半導体シーケンサーを用いた口腔扁平上皮癌の全エクソームシーケンス

田村 みゆき, 佐々木 泰史, 中垣 貴文, 小山 良太, 大箸 智子, 井戸川 雅史, 荻 和弘, 平塚 博義, 時野 隆至

日本癌学会総会記事 ( (一社)日本癌学会 )  75回   P - 1033  2016年10月

口腔扁平上皮癌におけるがん関連50遺伝子の変異解析

佐々木 泰史, 中垣 貴文, 田村 みゆき, 小山 良太, 福島 久代, 大箸 智子, 井戸川 雅史, 荻 和弘, 平塚 博義, 時野 隆至

日本癌学会総会記事 ( (一社)日本癌学会 )  75回   P - 1032  2016年10月

癌におけるp53および長鎖非コードRNA(IncRNA)による転写ネットワーク解析

井戸川 雅史, 大箸 智子, 佐々木 泰史, 時野 隆至

日本癌学会総会記事 ( (一社)日本癌学会 )  75回   J - 3026  2016年10月

がん関連遺伝子target sequenceによる食道扁平上皮癌の解析

岩谷 岳, 遠藤 史隆, 久米 浩平, 佐々木 泰史, 時野 隆至, 澤田 元太, 新井田 厚司, 三森 功士, 西塚 哲

日本癌学会総会記事 ( (一社)日本癌学会 )  75回   J - 2024  2016年10月

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網羅的解析

中垣 貴文, 佐々木 泰史, 井戸川 雅史, 小山 良太, 田村 みゆき, 大箸 智子, 荻 和弘, 平塚 博義, 時野 隆至

日本癌学会総会記事 ( (一社)日本癌学会 )  75回   P - 2297  2016年10月

次世代シークエンサーを用いた多発性骨髄腫における遺伝子変異とコピー数異常の解析

福島 久代, 佐々木 泰史, 田村 みゆき, 池田 博, 石黒 一也, 櫻井 晃洋, 時野 隆至

日本癌学会総会記事 ( (一社)日本癌学会 )  75回   P - 1024  2016年10月

多発性骨髄腫に有効なヒストンメチル化阻害薬の探索

石黒 一也, 北嶋 洋志, 新沼 猛, 丸山 玲緒, 甲斐 正広, 西山 廣陽, 進藤 哲哉, 池田 博, 石田 禎夫, 佐々木 泰史, 時野 隆至, 仲瀬 裕志, 鈴木 拓

日本癌学会総会記事 ( (一社)日本癌学会 )  75回   P - 2096  2016年10月