Misc - 小島　隆

肺腺癌におけるタイト結合分子cingulinおよび転写因子FOXO1の役割

石井 大智, 進藤 悠真, 新井 航, 金野 匠, 幸野 貴之, 宮島 正博, 小島 隆, 渡辺 敦

肺癌 ( (NPO)日本肺癌学会 )  62 ( 6 ) 635 - 635  2022年11月

正常ヒト肺上皮細胞におけるTGF-βとHDAC阻害剤のタイト結合分子および上皮バリアへの影響

進藤 悠真, 新井 航, 金野 匠, 幸野 貴之, 宮島 正博, 小島 隆, 渡辺 敦

日本呼吸器外科学会雑誌 ( (一社)日本呼吸器外科学会 )  35 ( 3 ) RO18 - 3  2021年05月

頭頸部扁平上皮癌の癌微小環境におけるAEBP1の解析

萬 顕, 山本 圭佑, 小幡 和史, 大國 毅, 黒瀬 誠, 近藤 敦, 高澤 啓, 小島 隆, 鈴木 拓, 高野 賢一

日本耳鼻咽喉科学会会報 ( (一社)日本耳鼻咽喉科頭頸部外科学会 )  124 ( 4 ) 690 - 690  2021年04月

タイト結合分子Cluadin-2をターゲットにした子宮内膜癌治療のin vitroにおける検討

嶋田 浩志, 岡田 匡氷, 金野 匠, 幸野 貴之, 松浦 基樹, 郷久 晴朗, 岩崎 雅宏, 小島 隆, 齋藤 豪

日本婦人科腫瘍学会雑誌 ( (公社)日本婦人科腫瘍学会 )  39 ( 1 ) 285 - 285  2021年01月

タイト結合による子宮内膜の悪性化制御機序

郷久 晴朗, 嶋田 浩志, 岩崎 雅宏, 染谷 真行, 金野 匠, 菊池 真, 幸野 貴之, 玉手 雅人, 松浦 基樹, 小島 隆, 齋藤 豪

北日本産科婦人科学会総会・学術講演会プログラム・抄録集 ( 東北連合産科婦人科学会・北日本産科婦人科学会 )  67回   36 - 36  2019年09月

子宮内膜悪性化におけるタイト結合分子claudin-2の関与

郷久 晴朗, 嶋田 浩志, 岩崎 雅宏, 秋元 太志, 西村 庸子, 玉手 雅人, 寺田 倫子, 松浦 基樹, 金野 匠, 幸野 貴之, 小島 隆, 齋藤 豪

日本婦人科腫瘍学会雑誌 ( (公社)日本婦人科腫瘍学会 )  37 ( 3 ) 497 - 497  2019年06月

子宮内膜癌細胞におけるタイト結合分子claudin-2の関与

郷久 晴朗, 嶋田 浩志, 玉手 雅人, 秋元 太志, 西村 庸子, 寺田 倫子, 松浦 基樹, 岩崎 雅宏, 小島 隆, 齋藤 豪

日本臨床細胞学会雑誌 ( (公社)日本臨床細胞学会 )  58 ( Suppl.1 ) 320 - 320  2019年05月

「アレルギーと環境の新しい考え方」に寄せる ヒト鼻粘膜上皮におけるtricellular Tight Junctionの形態・機能解析

大國 毅, 氷見 徹夫, 高野 賢一, 小島 隆

アレルギーの臨床 ( (株)北隆館 )  38 ( 7 ) 655 - 659  2018年07月

　概要を見る

鼻腔を含む気道上皮では、タイト結合により上皮細胞どうしが強固に接着・集合し、病原体の侵入を防ぐ上皮バリアが機能する。2細胞間のタイト結合ストランドが収束する3細胞間隙では、その中心にcentral tubeを形成する特殊な環境となり、これが樹状細胞の突起伸長経路、また物質透過の通路と推察される。今回われわれは、ヒト鼻粘膜上皮における3細胞間隙タイト結合の構造・調節機構を明らかにし、鼻粘膜を介したDrug delivery systemの可能性について検討した。(著者抄録)

タイト結合分子に焦点を当てた頭頸部扁平上皮癌におけるヒストン脱アセチル化酵素(HDAC)阻害剤の抗腫瘍作用機序の検討

垣内 晃人, 高野 賢一, 氷見 徹夫, 小島 隆

耳鼻咽喉科免疫アレルギー ( 日本耳鼻咽喉科免疫アレルギー学会 )  36 ( 2 ) 79 - 79  2018年06月

多機能な細胞接着分子が関わるがんの理解 診断・治療への応用をめざして 3細胞間タイト結合分子LSRの発現低下によるがん細胞の悪性化機序

金野 匠, 嶋田 浩志, 郷久 晴朗, 齋藤 豪, 幸野 貴之, 小島 隆

日本病理学会会誌 ( (一社)日本病理学会 )  107 ( 1 ) 242 - 242  2018年04月

ヒト鼻粘膜上皮細胞での上皮バリアおよび線毛形成の調節における転写因子p63の役割

金子 躍人, 幸野 貴之, 角木 拓也, 高野 賢一, 小笠原 徳子, 宮田 遼, 大國 毅, 矢島 諒人, 垣内 晃人, 小島 隆, 氷見 徹夫

日本耳鼻咽喉科学会会報 ( (一社)日本耳鼻咽喉科学会 )  121 ( 4 ) 567 - 567  2018年04月

ヒストン脱アセチル化酵素(HDAC)阻害剤による頭頸部扁平上皮癌タイト結合分子の変化と抗腫瘍効果

垣内 晃人, 角木 拓也, 黒瀬 誠, 高野 賢一, 近藤 敦, 小幡 和史, 野村 一顕, 宮田 遼, 金子 躍人, 氷見 徹夫, 小島 隆

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   [1P - 0386]  2017年12月

細胞質分裂における3細胞間タイト結合分子の局在と役割

金野 匠, 幸野 貴之, 菊池 真, 嶋田 浩志, 郷久 晴朗, 齋藤 豪, 小島 隆

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 - 0351]  2017年12月

3細胞間タイト結合蛋白LSRの頭頸部扁平上皮癌における発現と役割

垣内 晃人, 角木 拓也, 高野 賢一, 金子 躍人, 黒瀬 誠, 近藤 敦, 幸野 貴之, 氷見 徹夫, 小島 隆

耳鼻咽喉科免疫アレルギー ( 日本耳鼻咽喉科免疫アレルギー学会 )  35 ( 2 ) 102 - 102  2017年08月

3細胞間タイト結合分子LSRはAMOTに活性化されたHippo pathwayを介し子宮内膜癌の悪性化に関与する

郷久 晴朗, 嶋田 浩志, 阿部 秀悦, 玉手 雅人, 寺田 倫子, 鈴木 美和, 寺本 瑞絵, 秋元 太志, 野藤 五沙, 小川 万梨絵, 田中 綾一, 松浦 基樹, 岩崎 雅宏, 小島 隆, 齋藤 豪

日本婦人科腫瘍学会雑誌 ( (公社)日本婦人科腫瘍学会 )  35 ( 3 ) 527 - 527  2017年06月

がん細胞の上皮バリアおよび悪性化における糖代謝の役割

金野 匠, 幸野 貴之, 及能 大輔, 嶋田 浩志, 郷久 晴朗, 角木 拓也, 金子 躍人, 高野 賢一, 齋藤 豪, 小島 隆

日本細胞生物学会大会講演要旨集 ( (一社)日本細胞生物学会 )  69回   95 - 95  2017年05月

3細胞間タイト結合蛋白LSRはYAP/AMOT/merlinを介して子宮内膜癌の悪性化へ関与する

阿部 秀悦, 嶋田 浩志, 郷久 晴朗, 金野 匠, 齋藤 豪, 幸野 貴之, 小島 隆

日本細胞生物学会大会講演要旨集 ( (一社)日本細胞生物学会 )  69回   95 - 95  2017年05月

頭頸部扁平上皮癌におけるヒストン脱アセチル化酵素(HDAC)阻害剤のp63依存性タイト結合分子を介した抗腫瘍効果

垣内 晃人, 角木 拓也, 黒瀬 誠, 高野 賢一, 近藤 敦, 小幡 和史, 野村 一顕, 宮田 遼, 金子 躍人, 氷見 徹夫, 小島 隆

日本細胞生物学会大会講演要旨集 ( (一社)日本細胞生物学会 )  69回   95 - 95  2017年05月

ヒト鼻粘膜上皮バリアにおける転写制御因子p63の役割

金子 躍人, 角木 拓也, 高野 賢一, 小笠原 徳子, 横田 伸一, 氷見 徹夫, 幸野 貴之, 小島 隆

日本病理学会会誌 ( (一社)日本病理学会 )  106 ( 1 ) 321 - 321  2017年03月

膵癌の悪性化における3細胞間タイト結合分子LSRの役割

金野 匠, 及能 大輔, 河野 剛, 有元 千紘, 畠山 翔翼, 高橋 駿太, 幸野 貴之, 小島 隆

日本病理学会会誌 ( (一社)日本病理学会 )  106 ( 1 ) 293 - 293  2017年03月

3細胞間タイト結合蛋白LSRはHippo pathwayを介して子宮内膜癌の悪性化に関与している

嶋田 浩志, 郷久 晴朗, 金野 匠, 高橋 駿太, 畠山 翔翼, 幸野 貴之, 田中 綾一, 寺本 瑞絵, 岩崎 雅宏, 小島 隆, 齋藤 豪

日本婦人科腫瘍学会雑誌 ( (公社)日本婦人科腫瘍学会 )  34 ( 3 ) 416 - 416  2016年06月

ヒト鼻粘膜上皮バリアにおけるp63の役割

小島 隆, 角木 拓也, 金子 躍人, 高野 賢一, 小笠原 徳子, 横田 伸一, 氷見 徹夫, 幸野 貴之

日本細胞生物学会大会講演要旨集 ( (一社)日本細胞生物学会 )  68回   63 - 63  2016年05月

温度感受性マウス内耳有毛前駆細胞におけるMetforminおよびHDAC阻害剤による3細胞間タイト結合蛋白の誘導

角木 拓也, 金子 躍人, 高野 賢一, 氷見 徹夫, 幸野 貴之, 小島 隆

日本細胞生物学会大会講演要旨集 ( (一社)日本細胞生物学会 )  68回   99 - 99  2016年05月

頭頸部扁平上皮癌におけるp63/GATA-3経路を介したタイト結合分子JAM-Aの発現調節機構

角木 拓也, 黒瀬 誠, 高野 賢一, 近藤 敦, 小幡 和史, 野村 一顕, 宮田 遼, 金野 匠, 高橋 駿太, 畠山 翔翼, 幸野 貴之, 氷見 徹夫, 小島 隆

日本細胞生物学会大会講演要旨集 ( (一社)日本細胞生物学会 )  67回   134 - 134  2015年06月

頭頸部扁平上皮癌細胞株におけるp63を介したタイト結合分子JAM-Aの発現調節機構

高橋 駿太, 角木 拓也, 宮田 遼, 野村 一顕, 黒瀬 誠, 近藤 敦, 高野 賢一, 幸野 貴之, 氷見 徹夫, 小島 隆

日本細胞生物学会大会講演要旨集 ( (一社)日本細胞生物学会 )  66回   140 - 140  2014年05月

ROLE OF TIGHT JUNCTIONS IN SIGNAL TRANSDUCTION: AN UPDATE

Kenichi Takano, Takashi Kojima, Norimasa Sawada, Tetsuo Himi

EXCLI JOURNAL ( EXCLI JOURNAL MANAGING OFFICE )  13   1145 - 1162  2014年

書評論文，書評，文献紹介等  

　概要を見る

Tight junctions (TJs), which are the most apically located of the intercellular junctional complexes, have a barrier function and a fence function. Recent studies show that they also participate in signal transduction mechanisms. TJs are modulated by intracellular signaling pathways including protein kinase C, mitogen-activated protein kinase, and NF-kappa B, to affect the epithelial barrier function in response to diverse stimuli. TJs are also regulated by various cytokines, growth factors, and hormones via signaling pathways. To investigate the regulation of TJ molecules via signaling pathways in human epithelial cells under normal and pathological conditions, we established a novel model of human telomerase reverse transcriptase-transfected human epithelial cells. In this review, we describe the recent progress in our understanding of the role of TJs for signal transduction under normal conditions in upper airway epithelium, pancreatic duct epithelial cells, hepatocytes, and endometrial epithelial cells, and in pathological conditions including cancer and infection.

膵癌の上皮間葉移行におけるタイト結合分子claudin‐1に対するPKCα阻害剤の効果

及能大輔, 小島隆, 河野剛, 山口洋志, 伊東竜哉, 今村将史, 木村康利, 澤田典均, 平田公一

日本消化器癌発生学会総会プログラム・抄録集   25th   86  2014年

J-GLOBAL

Expression of tricellulin in epithelial cells and non-epithelial cells

Takashi Kojima, Takafumi Ninomiya, Takumi Konno, Takayuki Kohno, Masahiko Taniguchi, Norimasa Sawada

HISTOLOGY AND HISTOPATHOLOGY ( F HERNANDEZ )  28 ( 11 ) 1383 - 1392  2013年11月

書評論文，書評，文献紹介等  

　概要を見る

Tricellulin is the first molecular component of tricellular tight junctions at tricellular contacts where three epithelial cells meet, and it is required for the their formation and maintenance of the epithelial barrier. Tricellulin binds other tight junction proteins, and its expression and distribution are affected by the bicellular tight junction protein occludin and lipolysis-stimulated lipoprotein receptor (LSR) which is expressed at tricellular contacts. Tricellulin is also detected in endothelial cells, neurons, microglia and astrocytes. Here, we focused tricellulin expression in various types of epithelial cells, nasal epithelial cells, pancreatic duct epithelial cells cells and hepatocytes, and non-epithelial cells, dendritic cells and Schwann cells, compared to expression of the bicellular tight junction protein occludin and LSR, and discuss the regulation and the role of tricellulin in cellular specificity.

膵癌組織および膵癌細胞株におけるタイト結合分子tricellulinの核への局在

村田 雅樹, 高澤 啓, 田中 敏, 小島 隆, 澤田 典均

日本病理学会会誌 ( (一社)日本病理学会 )  102 ( 1 ) 473 - 473  2013年04月

胆道癌におけるclaudin18、maspin、p53の免疫組織化学的検討

計良 淑子, 高澤 啓, 村田 雅樹, 荻野 次郎, 及能 大輔, 中西 勝也, 田中 敏, 小島 隆, 長谷川 匡, 澤田 典均

日本病理学会会誌 ( (一社)日本病理学会 )  102 ( 1 ) 471 - 471  2013年04月

PKCα阻害剤は膵癌細胞のEMTにおけるタイト結合分子claudin‐1の低下を抑制する

及能大輔, 小島隆, 山口洋志, 伊東竜哉, 信岡隆幸, 今村将史, 木村康利, 澤田典均, 平田公一

日本外科学会雑誌   114   683  2013年03月

J-GLOBAL

Regulation of Tight Junctions in Upper Airway Epithelium

Takashi Kojima, Mitsuru Go, Ken-ichi Takano, Makoto Kurose, Tsuyoshi Ohkuni, Jun-ichi Koizumi, Ryuta Kamekura, Noriko Ogasawara, Tomoyuki Masaki, Jun Fuchimoto, Kazufumi Obata, Satoshi Hirakawa, Kazuaki Nomura, Takashi Keira, Ryou Miyata, Nobuhiro Fujii, Hiroyuki Tsutsumi, Tetsuo Himi, Norimasa Sawada

BIOMED RESEARCH INTERNATIONAL ( HINDAWI PUBLISHING CORPORATION )   2013年

書評論文，書評，文献紹介等  

　概要を見る

The mucosal barrier of the upper respiratory tract including the nasal cavity, which is the first site of exposure to inhaled antigens, plays an important role in host defense in terms of innate immunity and is regulated in large part by tight junctions of epithelial cells. Tight junction molecules are expressed in both M cells and dendritic cells as well as epithelial cells of upper airway. Various antigens are sampled, transported, and released to lymphocytes through the cells in nasal mucosa while they maintain the integrity of the barrier. Expression of tight junction molecules and the barrier function in normal human nasal epithelial cells (HNECs) are affected by various stimuli including growth factor, TLR ligand, and cytokine. In addition, epithelial-derived thymic stromal lymphopoietin (TSLP), which is a master switch for allergic inflammatory diseases including allergic rhinitis, enhances the barrier function together with an increase of tight junction molecules in HNECs. Furthermore, respiratory syncytial virus infection in HNECs in vitro induces expression of tight junction molecules and the barrier function together with proinflammatory cytokine release.. is paper summarizes the recent progress in our understanding of the regulation of tight junctions in the upper airway epithelium under normal, allergic, and RSV-infected conditions.

DOI

扁桃・アデノイドはなぜあるのか？鼻はなにをしているのか？―小児の粘膜免疫・粘膜防御最前線―

氷見徹夫, 高野賢一, 山下恵司, 小笠原徳子, 正木智之, 小幡和史, 堤裕幸, 小島隆, 一宮慎吾, 澤田典均, 横田伸一

小児耳鼻咽喉科   34   239 - 244  2013年

DOI

ウイルス性上気道感染での免疫応答と鼻粘膜上皮の役割

氷見徹夫, 高野賢一, 大國毅, 小笠原徳子, 正木智之, 小幡和史, 堤裕幸, 小島隆, 澤田典均, 横田伸一

耳鼻咽喉科展望   56   162 - 177  2013年  [招待有り]

記事・総説・解説・論説等（その他）  

DOI

Regulation of Claudin-4 via Protein Kinase C Signal Pathway in Normal Human Pancreatic Duct Epithelial Cells and Cancer Cells

H. Yamaguchi, T. Kojima, D. Kyuno, T. Ito, M. Imamura, Y. Kimura, N. Sawada, K. Hirata

PANCREAS ( LIPPINCOTT WILLIAMS & WILKINS )  41 ( 7 ) 1149 - 1149  2012年10月

研究発表ペーパー・要旨（国際会議）  

Protein Kinase C-alpha Regulates Claudin-1 via Snail-dependent and -independent Pathways in Pancreatic Cancer

D. Kyuno, T. Kojima, H. Yamaguchi, T. Ito, M. Imamura, Y. Kimura, A. Takasawa, M. Murata, M. Murata, K. Hirata

PANCREAS ( LIPPINCOTT WILLIAMS & WILKINS )  41 ( 7 ) 1150 - 1150  2012年10月

研究発表ペーパー・要旨（国際会議）  

PKCαによるSnailを介した膵癌のタイト結合分子への影響(PKC-alpha regulates claudin-1 via Snail gene in pancreatic cancer)

及能 大輔, 小島 隆, 山口 洋志, 伊東 竜哉, 今村 将史, 木村 康利, 高澤 啓, 村田 雅樹, 平田 公一, 澤田 典均

日本癌学会総会記事 ( 日本癌学会 )  71回   295 - 295  2012年08月

タイト結合分子tricellulinの核における発現はヒト膵癌細胞において細胞増殖に関与している(Expression of tricellulin in the nuclei is associated with cell proliferation in human pancreatic cancer cells)

村田 雅樹, 高澤 啓, 小島 隆, 澤田 典均

日本癌学会総会記事 ( (一社)日本癌学会 )  71回   525 - 525  2012年08月

胆道癌におけるclaudin-18、Maspinの免疫組織化学的検討(Immunohistochemical investigation of claudin-18 and Maspin in biliary tract cancer)

計良 淑子, 高澤 啓, 村田 雅樹, 小島 隆, 長谷川 匡, 澤田 典均

日本癌学会総会記事 ( (一社)日本癌学会 )  71回   351 - 351  2012年08月

ヒト膵管上皮細胞株におけるJun N-terminal kinase阻害剤によるバリア機能の亢進

金野 匠, 小島 隆, 及能 大輔, 村田 雅樹, 高澤 啓, 田中 敏, 澤田 典均

日本病理学会会誌 ( (一社)日本病理学会 )  101 ( 1 ) 438 - 438  2012年03月

PKCα阻害剤による膵癌細胞株および正常ヒト膵管上皮細胞のタイト結合分子への影響

及能 大輔, 小島 隆, 伊東 竜哉, 山口 洋志, 高澤 啓, 澤田 典均

日本病理学会会誌 ( (一社)日本病理学会 )  101 ( 1 ) 264 - 264  2012年03月

ヒト鼻粘膜上皮細胞を用いたcurcuminによるRSvirusの複製阻害およびタイト結合発現誘導のメカニズム解析

小幡 和史, 小島 隆, 正木 智之, 渕本 潤, 平川 賢史, 野村 一顕, 高澤 啓, 小笠原 徳子, 氷見 徹夫, 澤田 典均

日本病理学会会誌 ( (一社)日本病理学会 )  101 ( 1 ) 280 - 280  2012年03月

ヒト鼻粘膜上皮細胞系におけるrespiratory syncytial virus感染によるMatrix Metalloproteinaseの増加

平川 賢史, 小島 隆, 小幡 和史, 野村 一顕, 正木 智之, 渕本 潤, 高澤 啓, 堤 裕幸, 澤田 典均

日本病理学会会誌 ( (一社)日本病理学会 )  101 ( 1 ) 327 - 327  2012年03月

肝炎および肝癌におけるタイト結合分子の発現変化

辻脇 光洋, 村田 雅樹, 高澤 啓, 平塚 佑太郎, 福田 理英子, 田中 敏, 小島 隆, 澤田 典均

日本病理学会会誌 ( (一社)日本病理学会 )  101 ( 1 ) 360 - 360  2012年03月

胆道癌におけるclaudin-18、maspinの免疫組織化学的検討

高澤 啓, 計良 淑子, 村田 雅樹, 荻野 次郎, 及能 大輔, 田中 敏, 小島 隆, 長谷川 匡, 澤田 典均

日本病理学会会誌 ( (一社)日本病理学会 )  101 ( 1 ) 288 - 288  2012年03月

膵癌組織および膵癌細胞株におけるタイト結合分子tricellulinの核への局在

村田 雅樹, 小野 佑輔, 高澤 啓, 田中 敏, 小島 隆, 澤田 典均

日本病理学会会誌 ( (一社)日本病理学会 )  101 ( 1 ) 264 - 264  2012年03月

緑膿菌産生エラスターゼによるヒト鼻粘膜上皮バリアへの影響

野村 一顕, 小島 隆, 小幡 和史, 平川 賢史, 高澤 啓, 氷見 徹夫, 澤田 典均

日本病理学会会誌 ( (一社)日本病理学会 )  101 ( 1 ) 327 - 327  2012年03月

P2-072  アトピー性皮膚炎の表皮におけるp63依存性TSLP-TSLP受容体経路

久保 輝文, 叶 汭, 三橋 由佳梨, 山下 恵司, 佐藤 明紀, 小島 隆, 佐藤 昇志, 一宮 慎吾

日本臨床免疫学会会誌 ( 日本臨床免疫学会 )  35 ( 4 ) 362b - 362b  2012年

　概要を見る

アトピー性皮膚炎は本邦の小児の約1割が罹患し，罹患者の生活の質を著しく低下させる．さらにアトピー性皮膚炎自体が気管支喘息やアレルギー性結膜炎など種々の慢性アレルギー疾患の根底にある可能性が分子免疫学的にも支持されつつあり，その病態の機序の解明が急がれている．   近年，アトピー性皮膚炎の患部のケラチノサイトがthymic stromal lymphopoietin（TSLP）を高産生することが明らかとなった．TSLPはTSLP受容体（TSLPR）を発現する樹状細胞を介してTh0細胞をTh2細胞へと分化させることで，アトピー・アレルギー疾患の病態形成に中心的役割を果たすと考えられている．   我々は今回，表皮ケラチノサイトにおけるTSLPRの発現を明らかとした．また，表皮ケラチノサイト初代培養細胞および表皮モデルHaCaT細胞を用いた検討において，TSLPRの発現は表皮幹細胞因子p63を介した自然免疫経路によって制御されていた．興味深いことに表皮に対するTSLP刺激は更なるTSLP産生を惹起し，その他の炎症性サイトカインの発現も促進していた．これらの結果からTSLPはオートクラインあるいはパラクラインによってケラチノサイト自身にも作用するループを形成し，アトピー性皮膚炎患部表皮の炎症環境の形成，維持に関与していると考えられる．

DOI CiNii

S22-4 鼻粘膜上皮のバリア機能と免疫調節機構としての位置づけ(S22 chronic rhinosinusitisの免疫,シンポジウム,第61回日本アレルギー学会秋季学術大会)

氷見 徹夫, 高野 賢一, 郷 充, 大國 毅, 亀倉 隆太, 小幡 和史, 計良 宗, 小島 隆, 澤田 典均

アレルギー ( 一般社団法人日本アレルギー学会 )  60 ( 9 )  2011年10月

CiNii

膵癌におけるタイト結合分子Claudin-18のPKC経路およびDNAメチル化を介した発現調節機構

伊東 竜哉, 及能 大輔, 山口 洋志, 小島 隆, 目黒 誠, 今村 将史, 木村 康利, 高澤 啓, 村田 雅樹, 澤田 典均, 平田 公一

日本消化器病学会雑誌 ( (一財)日本消化器病学会 )  108 ( 臨増大会 ) A902 - A902  2011年09月

膵切除標本を利用したヒト膵管上皮細胞培養系の確立と膵管上皮タイト結合蛋白解析

山口 洋志, 小島 隆, 伊東 竜哉, 及能 大輔, 木村 康利, 今村 将史, 永山 稔, 澤田 典均, 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 ) 350 - 350  2011年05月

上皮産生TSLPのヒト鼻粘膜における役割

亀倉 隆太, 小島 隆, 高野 賢一, 黒瀬 誠, 郷 充, 澤田 典均, 氷見 徹夫

日本鼻科学会会誌 ( (一社)日本鼻科学会 )  50 ( 1 ) 38 - 45  2011年04月

　概要を見る

胸腺間質性リンパ球新生因子(TSLP)の鼻粘膜(NM)上皮における役割を解明するために、アレルギー性鼻炎(AR)患者のNMにおけるTSLPの発現と上皮内樹状細胞の活性化との関係、NM上皮産生TSLPの発現誘導のメカニズム、TSLPによる培養NM上皮細胞および樹状細胞のタイト結合蛋白への影響について検討した。その結果、上皮構造が比較的保たれている正常またはAR発症の初期段階では、NM上皮から産生されたTSLPはNM上皮や樹状細胞のタイト結合の発現を増強する働きを有しており、NM上皮自らのバリア機能を維持することで鼻腔内の抗原が無秩序に上皮下に侵入することを防ぎ、樹状細胞による選択的な抗原取り込みを助けるという、TSLPによるNM特有の自然免疫を介した生体防御機構が存在することが示唆された。

膵癌細胞株及び正常膵管上皮細胞におけるタイト結合とPKCαとの関係

及能 大輔, 小島 隆, 伊東 竜哉, 山田 洋志, 辻脇 光洋, 高澤 啓, 村田 雅樹, 田中 敏, 澤田 典均

日本病理学会会誌 ( (一社)日本病理学会 )  100 ( 1 ) 319 - 319  2011年03月

卵巣癌細胞株におけるタイト結合分子claudin4に対するCPE作用条件についての検討

井上 万梨絵, 田中 敏, 染谷 真行, 高澤 啓, 村田 雅樹, 小島 隆, 斎藤 豪, 澤田 典均

日本病理学会会誌 ( (一社)日本病理学会 )  100 ( 1 ) 459 - 459  2011年03月

培養ヒト膵管上皮細胞におけるtricellulin発現調節機構の解明 Caスイッチによる発現・局在変化を中心に

高澤 啓, 小島 隆, 及能 大輔, 辻脇 光洋, 村田 雅樹, 田中 敏, 澤田 典均

日本病理学会会誌 ( (一社)日本病理学会 )  100 ( 1 ) 318 - 318  2011年03月

Expression and Function of Tight Junctions in the Crypt Epithelium of Human Palatine Tonsils

Mitsuru Go, Takashi Kojima, Ken-ichi Takano, Makoto Kurose, Tsuyoshi Ohkuni, Tomoyuki Masaki, Ryuta Kamekura, Atsushi Kondo, Noriko Ogasawara, Tetsuo Himi, Norimasa Sawada

RECENT ADVANCES IN TONSILS AND MUCOSAL BARRIERS OF THE UPPER AIRWAYS ( KARGER )  72   185 - 186  2011年

研究発表ペーパー・要旨（国際会議）  

Respiratory Syncytial Virus Infection and the Tight Junctions of Nasal Epithelial Cells

Hiroyuki Tsutsumi, Takashi Kojima, Satoshi Hirakawa, Tomoyuki Masaki, Tamaki Okabayashi, Shinichi Yokota, Nobuhiro Fujii, Tetsuo Himi, Norimasa Sawada

RECENT ADVANCES IN TONSILS AND MUCOSAL BARRIERS OF THE UPPER AIRWAYS ( KARGER )  72   153 - 156  2011年  [招待有り]

記事・総説・解説・論説等（国際会議プロシーディングズ）  

　概要を見る

Respiratory syncytial virus (RSV) primarily infects upper respiratory tract cells, mainly nasal epithelial cells. The tight junctions of nasal epithelial cells are thought to perform important innate immune function against foreign materials including respiratory viruses. We investigated in vitro the relationship of RSV infection and the tight junctions of primary nasal epithelial cells which had been transfected with human telomerase reverse transcriptase (hTERT) to prolong cell life. Nasal epithelial cells developed tight junctions when cultured in medium containing fetal bovine serum, and these cells showed apparent resistance to RSV infection compared to control cells. RSV could infect these cells from apical but not basolateral side, suggesting that only apical side possess RSV receptor or a mechanism for absorbing RSV particles. Importantly, RSV infection of the cells enhanced the expression of tight junction proteins occludin, claudin-4 and ZO-1. These findings suggest that RSV infection induces polarity in the infected cells. This polarity could facilitate cellular secretion of propagated RSV, thereby spreading the infection. Copyright (C) 2011 S. Karger AG, Basel

DOI

MW1-5 鼻粘膜上皮におけるIL-33の役割(MW1 アレルギー性鼻炎・結膜炎研究の進歩,ミニワークショップ,第60回日本アレルギー学会秋季学術大会)

亀倉 隆太, 高野 賢一, 郷 充, 小島 隆, 澤田 典均, 氷見 徹夫

アレルギー ( 一般社団法人日本アレルギー学会 )  59 ( 9 )  2010年10月

CiNii

RSウイルス感染ヒト鼻粘膜上皮細胞におけるタイト結合の誘導

正木 智之, 小島 隆, 大國 毅, 小笠原 徳子, 岡林 環樹, 澤田 典均, 氷見 徹夫

耳鼻咽喉科免疫アレルギー ( 日本耳鼻咽喉科免疫アレルギー学会 )  28 ( 2 ) 81 - 82  2010年09月

膵癌細胞株におけるタイト結合蛋白claudin-4、-18発現調節機構の解析

伊東 竜哉, 小島 隆, 山口 洋志, 木村 康利, 永山 稔, 今村 将史, 澤田 典均, 平田 公一

日本消化器外科学会総会 ( (一社)日本消化器外科学会 )  65回   150 - 150  2010年07月

ヒト鼻粘膜上皮細胞における新規タイト結合分子tricellulinの発現調節

大國 毅, 小島 隆, 小笠原 徳子, 正木 智之, 澤田 典均, 氷見 徹夫

日本鼻科学会会誌 ( (一社)日本鼻科学会 )  49 ( 3 ) 425 - 425  2010年07月

What’s New in SURGERY FRONTIER 第65回 消化器癌と細胞間接着 4)消化器癌とタイト結合・クローディン

伊東竜哉, 山口洋志, 小島隆, 澤田典均, 平田公一

Surg Front   17 ( 2 ) 158-161,106  2010年06月

J-GLOBAL

Toll様受容体刺激によるヒト鼻粘膜上皮細胞タイト結合分子発現変化について

大國 毅, 小島 隆, 小笠原 徳子, 正木 智之, 澤田 典均, 氷見 徹夫

日本耳鼻咽喉科学会会報 ( (一社)日本耳鼻咽喉科頭頸部外科学会 )  113 ( 4 ) 405 - 405  2010年04月

膵癌のタイト結合分子をターゲットとした新規治療法開発に向けての基礎的研究

伊東竜哉, 小島隆, 山口洋志, 木村康利, 永山稔, 今村将史, 秋月恵美, 澤田典均, 平田公一

日本外科学会雑誌   111   394  2010年03月

J-GLOBAL

新規タイト結合蛋白tricellulinのJun N-terminal kinase(JNK)経路を介した発現調節機構の解明

小島 隆, 村田 雅樹, 田中 敏, 澤田 典均

日本病理学会会誌 ( (一社)日本病理学会 )  99 ( 1 ) 225 - 225  2010年03月

RSウイルス感染ヒト鼻粘膜上皮細胞におけるタイト結合の誘導

正木 智之, 小島 隆, 大國 毅, 小笠原 徳子, 氷見 徹夫, 村田 雅樹, 田中 敏, 澤田 典均

日本病理学会会誌 ( (一社)日本病理学会 )  99 ( 1 ) 191 - 191  2010年03月

鼻粘膜上皮細胞における自然免疫toll like receptorによるタイト結合分子発現調節

大國 毅, 小島 隆, 小笠原 徳子, 正木 智之, 澤田 均典, 氷見 徹夫

アレルギーの臨床 ( (株)北隆館 )  30 ( 1 ) 88 - 89  2010年01月

膵癌細胞株のタイト結合におけるPKCalphaの役割

及能大輔, 小島隆, 伊東竜哉, 山口洋志, 平田公一, 澤田典均

日本臨床分子形態学会総会・学術集会講演プログラム・要旨集   42nd   102  2010年

J-GLOBAL

新規培養ヒト正常膵管上皮細胞を用いたタイト結合蛋白claudin‐4を標的とした膵癌分子標的治療の基礎的研究

小島隆, 山口洋志, 伊東竜哉, 及能大輔, 平田公一, 澤田典均

日本臨床分子形態学会総会・学術集会講演プログラム・要旨集   42nd   42  2010年

J-GLOBAL

ヒト鼻粘膜上皮細胞を用いたRSウイルス感染モデルの確立と上皮タイト結合への影響

正木 智之, 小島 隆, 大國 毅, 小笠原 徳子, 岡林 環樹, 藤井 暢弘, 堤 裕幸, 澤田 典均, 氷見 徹夫

日本鼻科学会会誌 ( (一社)日本鼻科学会 )  48 ( 3 ) 323 - 323  2009年08月

ヒト鼻粘膜の機能解析から見えてくること

氷見 徹夫, 郷 充, 近藤 敦, 高野 賢一, 正木 智之, 小泉 純一, 亀倉 隆太, 大國 毅, 小笠原 徳子, 小島 隆, 澤田 典均

耳鼻咽喉科展望 ( 耳鼻咽喉科展望会 )  52 ( 補冊1 ) 9 - 18  2009年08月

　概要を見る

ヒト鼻粘膜上皮の初代培養系を用いたバリア機能の解析は経鼻投与による炎症制御の方法論の確立に重要である。タイト結合はバリア機能に関連し物質の透過性に関与する因子のひとつであり、この機能を調節するメカニズムを解明することは炎症制御に重要である。さらに、樹状細胞やM細胞(micro-folds cell)のような抗原サンプリングに関与する免疫担当細胞の検討は経鼻投与によるアレルギー・感染などの免疫療法の効率化に寄与する。また、タイト結合などの細胞接着に関与する因子はウイルスの受容体であることが示されており、この意味ではウイルス感染の予防のための治療戦略としてタイト結合タンパクの研究を進めることは意義のあることと考えられる。ここでは鼻粘膜上皮と上皮内樹状細胞、さらに鼻粘膜のM細胞の存在について言及し、タイト結合とウイルス感染との関連、さらに、タイト結合タンパクの調節のメカニズムについても言及する。(著者抄録)

ヒト膵管上皮細胞培養系の確立とタイト結合蛋白claudinを標的とした膵癌治療基礎研究

山口洋志, 小島隆, 伊東竜哉, 木村康利, 澤田典均, 平田公一

すい臓   24 ( 3 ) 372  2009年06月

J-GLOBAL

疾患モデル動物による検討 肝癌自然発症マウスでの知見--Cx32欠損マウスを用いて (肝癌--基礎・臨床研究のアップデート) -- (肝癌の発生・進展機序)

小島 隆, 村田 雅樹, 澤田 典均

日本臨床 ( 日本臨床社 )  67 ( 0 ) 105 - 109  2009年05月

CiNii

ヒト鼻粘膜上皮細胞と樹状細胞のタイト結合蛋白はthymic stromal lymphopoietin(TSLP)によって誘導される

亀倉 隆太, 小島 隆, 小笠原 徳子, 大國 毅, 高野 賢一, 郷 充, 澤田 典均, 氷見 徹夫

日本耳鼻咽喉科学会会報 ( (一社)日本耳鼻咽喉科頭頸部外科学会 )  112 ( 4 ) 376 - 376  2009年04月

膵癌細胞株のPKCシグナル異常におけるタイト結合蛋白の変化

伊東竜哉, 小島隆, 山口洋志, 村田雅樹, 田中敏, 千葉英樹, 平田公一, 澤田典均

日本病理学会会誌   98 ( 1 ) 255  2009年03月

J-GLOBAL

核内受容体PPAR-γを介したヒト鼻粘膜におけるタイト結合の発現調節機構

小笠原 徳子, 小島 隆, 郷 充, 亀倉 隆太, 大國 毅, 正木 智之, 村田 雅樹, 田中 敏, 千葉 英樹, 氷見 徹夫, 澤田 典均

日本病理学会会誌 ( (一社)日本病理学会 )  98 ( 1 ) 323 - 323  2009年03月

核内受容体PPARγを介したヒト鼻粘膜上皮細胞, 樹状細胞のタイト結合の調節機構 : アレルギー性鼻炎の予防・治療のための基礎研究

小笠原 徳子, 小島 隆, 郷 充, 亀倉 隆太, 高野 賢一, 大国 毅, 正木 智之, 澤田 典均, 氷見 徹夫

耳鼻咽喉科免疫アレルギー   27 ( 2 ) 82 - 83  2009年02月

CiNii

MS3-2 核内受容体PPAR-gammaを介したヒト鼻粘膜上皮細胞および樹状細胞のタイト結合の発現調節機構(抗原提示細胞,第58回日本アレルギー学会秋季学術大会)

小笠原 徳子, 小島 隆, 郷 充, 亀倉 隆太, 高野 賢一, 大國 毅, 澤田 典均, 氷見 徹夫

アレルギー ( 一般社団法人日本アレルギー学会 )  57 ( 9 )  2008年10月

CiNii

Drug Delivery System(DDS)としてのエアロゾルの将来性 鼻粘膜を介した新たな治療戦略の開発と応用 粘膜のバリアと抗原サンプリング 鼻粘膜を介した新たな治療戦略にむけて

郷 充, 小島 隆, 亀倉 隆太, 小笠原 徳子, 小泉 純一, 黒瀬 誠, 高野 賢一, 澤田 典均, 氷見 徹夫

耳鼻咽喉科展望 ( 耳鼻咽喉科展望会 )  51 ( 補冊1 ) 32 - 38  2008年08月

　概要を見る

近年、粘膜上皮の自然免疫の一つとして、バリア機能を有するタイト結合形成の関与が注目されている。我々はこれまで、ヒト鼻粘膜上皮においても発達したタイト結合による上皮バリアが存在し、一部のアレルギー性鼻炎においては、抗原提示細胞である樹状細胞が上皮と同様のタイト結合分子を発現し、鼻腔内の抗原を感知すべく樹状突起を伸ばしている特殊な機構があることを示した。さらに安定した培養が可能であるテロメラーゼ遺伝子導入ヒト鼻粘膜上皮細胞を用いて、炎症に関与がみられる増殖因子、サイトカインおよびTLRリガンドを処置し、タイト結合蛋白の発現およびバリア機能の変化について検索した。これらの結果は、タイト結合によるバリアを選択的あるいは効果的に制御することにより、鼻粘膜を介した新しい薬物送達の可能性があることを示している。(著者抄録)

ヒト鼻粘膜上皮バリアにおけるPKCシグナルを介したタイト結合の調節機構

小島 隆, 小泉 純一, 小笠原 徳子, 亀倉 隆太, 黒瀬 誠, 今野 信宏, 郷 充, 高野 賢一, 氷見 徹夫, 澤田 典均

Inflammation and Regeneration ( (一社)日本炎症・再生医学会 )  28 ( 4 ) 354 - 354  2008年07月

培養ヒト膵管上皮細胞におけるタイト結合蛋白claudinの解析とその展望

山口 洋志, 小島 隆, 木村 康利, 孫 誠一, 今村 将史, 永山 稔, 信岡 隆幸, 澤田 典均, 平田 公一

膵臓 ( (一社)日本膵臓学会 )  23 ( 3 ) 374 - 374  2008年06月

DP-124-1 肝タイト結合蛋白claudin-2の発現低下は毛細胆管形成を抑制する(第108回日本外科学会定期学術集会)

孫 誠一, 小島 隆, 今村 将史, 山口 洋志, 村田 雅樹, 澤田 典均, 平田 公一

日本外科学会雑誌 ( 一般社団法人日本外科学会 )  109 ( 2 )  2008年04月

CiNii

What's New in SURGERY FRONTIER 細胞間接着装置と消化器癌 タイト結合蛋白(occludin)と消化器癌

今村 将史, 小島 隆, 徳永 祐一, 平田 公一, 澤田 典均

Surgery Frontier ( (株)メディカルレビュー社 )  15 ( 1 ) 70 - 74  2008年03月

培養ヒト膵管上皮細胞におけるタイト結合蛋白Claudin4の発現調節機構

山口 洋志, 小島 隆, 孫 誠一, 村田 雅樹, 田中 敏, 千葉 英樹, 木村 康利, 平田 公一, 澤田 典均

日本病理学会会誌 ( (一社)日本病理学会 )  97 ( 1 ) 256 - 256  2008年03月

アレルギー性鼻炎における鼻粘膜上皮バリアと免疫調節機構

氷見 徹夫, 郷 充, 高野 賢一, 黒瀬 誠, 小泉 純一, 亀倉 隆太, 小笠原 徳子, 小島 隆, 澤田 典均

アレルギーの臨床 ( (株)北隆館 )  27 ( 13 ) 1038 - 1043  2007年12月

　概要を見る

鼻粘膜上皮は機械的バリアだけでなく、「粘膜免疫機構」という自然免疫・獲得免疫を成立させるための重要な「免疫臓器」であることが次々と明らかとなっている。鼻粘膜においてもタイト結合と樹状細胞との関係は、古典的な抗原感知・移送という概念とは異なる新しい抗原サンプリング機構のあることがわかった。さらに、アレルギー性鼻炎では、アレルギー発症に関与する重要なサイトカインであるTSLPが主に上皮に強く発現していることが確認され、鼻粘膜上皮がアレルギー調節機構を持つ細胞群として重要であることが考えられた。(著者抄録)

アレルギー性鼻炎におけるThymic stromal lymphopoietin(TSLP)の発現と上皮バリア機能への影響

亀倉 隆太, 小島 隆, 黒瀬 誠, 小泉 純一, 高野 賢一, 今野 信宏, 播摩谷 敦, 郷 充, 澤田 典均, 氷見 徹夫

耳鼻咽喉科免疫アレルギー ( 日本耳鼻咽喉科免疫アレルギー学会 )  25 ( 2 ) 63 - 64  2007年09月

　概要を見る

アレルギー性鼻炎患者の鼻粘膜組織を用いて、thymic stromal lymphopoietin(TSLP)の発現と上皮内樹状細胞の活性化との関係を検討した。また培養ヒト鼻粘膜上皮細胞を用いて、TSLPのタイト結合への影響を検討した。RT-PCRおよびWestern blot法を用いてTSLPのmRNAおよび蛋白レベルの発現をみると、アレルギー性鼻炎の鼻粘膜では正常鼻粘膜に比し、TSLPの発現がmRNAおよび蛋白レベルで亢進していた。また鼻粘膜上皮をTSLP高発現群と低発現群に分類して免疫染色を行い、上皮内に存在するLCA、CD11c陽性細胞数を計測すると、TSLP高発現群では低発現群に比し、上皮内にLCAおよびCD11c陽性細胞が多数認められた。TSLP処理した培養鼻粘膜上皮細胞では、濃度依存性にclaudin-1、-7、occludinの発現増加がみられ、transepithelial resistanceの増加も認められた。培養鼻粘膜上皮細胞を用いたTSLPが鼻粘膜上皮細胞自らのバリア機能を維持し、樹状細胞による選択的取り込みを助けている可能性を示唆した。アレルギー性鼻炎における鼻粘膜上皮によるTSLPの産生とタイト結合の機能亢進は、アレルギー性鼻炎の新しい診断および治療に繋がると考えられた。

ヒト鼻粘膜上皮における自然免疫に関与するタイト結合の調節機構

小泉 純一, 小島 隆, 播摩谷 敦, 今野 信宏, 黒瀬 誠, 高野 賢一, 亀倉 隆太, 郷 充, 澤田 典均, 氷見 徹夫

耳鼻咽喉科免疫アレルギー ( 日本耳鼻咽喉科免疫アレルギー学会 )  25 ( 2 ) 65 - 66  2007年09月

　概要を見る

初代培養ヒト鼻粘膜上皮細胞を用いて、12-O-tetradecanoylphorbol-13-acetate(TPA)処置によるタイト結合の発現と機能の変化を検索し、ヒト鼻粘膜上皮細胞のタイト結合の調節機構を検討した。バリア機能の変化をみると、TPA処置後のヒト鼻粘膜上皮細胞(TPA処置群)では7時間をピークとして、コントロール群に比し最高8倍の上昇が認められた。Permeabilityの検討では、70kD dextranにおいて、TPA処置群ではコントロール群に比し透過性が明らかに低下していた。PKCシグナル分子およびタイト結合分子の発現変化を検討すると、PCKシグナル分子では、TPA処置後2時間においてPhospho-pan-PKC、phospho-MARCKs、phospho-MAPKが増加し、PKC familyの中でもδ、ε、θが増加していた。タイト結合分子では、TPA処置後2時間において、ウエスタンブロット法にてclaudin-1、occludin、ZO-1、-2蛋白の増加を、RT-PCR法にてclaudin-1、-4、occludin、JAM-1、E-cadherin、ZO-1、-2 mRNAの増加を認めた。ヒト鼻粘膜上皮の自然免疫においてもタイト結合は重要であり、様々な抗原の曝露によりPKCシグナル経路を介したバリア機能の亢進が起こっている可能性が示唆された。

ヒト鼻粘膜上皮バリアにおけるPKCシグナルを介したタイト結合の調節機構

小泉 純一, 小島 隆, 黒瀬 誠, 亀倉 隆太, 小笠原 徳子, 今野 信宏, 郷 充, 澤田 典均, 氷見 徹夫

日本鼻科学会会誌 ( (一社)日本鼻科学会 )  46 ( 3 ) 263 - 263  2007年08月

【肝小葉ゾーンからみた肝病態と臨床】肝小葉ゾーンとその解剖学的、機能的意義 肝細胞タイト結合、claudinと肝小葉

小島 隆, 村田 雅樹, 澤田 典均

肝胆膵 ( (株)アークメディア )  55 ( 1 ) 45 - 54  2007年07月

テロメラーゼ遺伝子導入培養ヒト鼻粘膜上皮細胞のタイト結合の発現と機能

小島 隆, 黒瀬 誠, 亀倉 隆太, 小泉 純一, 小笠原 徳子, 氷見 徹夫, 澤田 典均

Inflammation and Regeneration ( (一社)日本炎症・再生医学会 )  27 ( 4 ) 439 - 439  2007年07月

黄疸時におけるタイト結合蛋白claudin-2の役割

今村 将史, 小島 隆, 孫 誠一, 木村 康利, 澤田 典均, 平田 公一

日本外科学会雑誌 ( (一社)日本外科学会 )  108 ( 臨増2 ) 197 - 197  2007年03月

EVS9-1 アレルギー性鼻炎における鼻粘膜上皮バリアと抗原認識・提示機構(イブニングシンポジウム9 鼻アレルギー-病態解明と治療の進歩-,第56回日本アレルギー学会秋季学術大会)

氷見 徹夫, 高野 賢一, 黒瀬 誠, 小泉 純一, 亀倉 隆太, 小島 隆, 澤田 典均

アレルギー ( 一般社団法人日本アレルギー学会 )  55 ( 8 )  2006年09月

CiNii

in vivoおよびin vitroにおけるヒト鼻粘膜上皮細胞のM様細胞とタイト結合

小泉 純一, 小島 隆, 黒瀬 誠, 高野 賢一, 亀倉 隆太, 澤田 典均, 氷見 徹夫

耳鼻咽喉科免疫アレルギー ( 日本耳鼻咽喉科免疫アレルギー学会 )  24 ( 2 ) 179 - 179  2006年09月

タイト結合による上皮細胞極性維持にはoccludinの細胞外第2ループが重要である

徳永 祐一, 小島 隆, 飛岡 弘敏, 村田 雅樹, 小山内 誠, 千葉 英樹, 澤田 典均

日本癌学会総会記事 ( (一社)日本癌学会 )  65回   242 - 242  2006年09月

肝病態と細胞接着分子の新知見 黄疸とタイト結合蛋白claudin

小島 隆, 今村 将史, 孫 誠一, 村田 雅樹, 平田 公一, 澤田 典均

Surgery Frontier ( (株)メディカルレビュー社 )  13 ( 2 ) 176 - 180  2006年06月

肝細胞のバリア機能におけるタイト結合蛋白claudin-2の役割

今村 将史, 小島 隆, 孫 誠一, 村田 雅樹, 小山内 誠, 千葉 英樹, 平田 公一, 澤田 典均

日本病理学会会誌 ( (一社)日本病理学会 )  95 ( 1 ) 191 - 191  2006年04月

病態発生とタイト結合とギャップ結合の役割 黄疸と血液胆汁関門

小島 隆, 村田 雅樹, 小山内 誠, 千葉 英樹, 澤田 典均

日本病理学会会誌 ( (一社)日本病理学会 )  95 ( 1 ) 155 - 155  2006年04月

The significance of interferon-gamma-triggered internalization of tight-junction proteins in inflammatory bowel disease.

Hideki Chiba, Takashi Kojima, Makoto Osanai, Norimasa Sawada

Science's STKE : signal transduction knowledge environment   2006 ( 316 ) pe1  2006年

書評論文，書評，文献紹介等  

　概要を見る

Disruption of the epithelial barrier function of tight junctions by the proinflammatory cytokine interferon (IFN)-gamma plays a fundamental role in the pathogenesis of inflammatory bowel disease and other gastrointestinal disorders, but its precise mechanism has not been established. This Perspective provides an overview of how IFN-gamma triggers barrier dysfunction and bacterial translocation in intestinal epithelial cells, and highlights the roles of internalization of tight-junction transmembrane proteins, as well as rearrangements of the cortical actin cytoskeleton, in intestinal inflammation.

DOI PubMed

タイト結合蛋白occludinは肝細胞において生存シグナルの伝達に重要な役割を有する

村田 雅樹, 小島 隆, 高野 賢一, 畠山 尚子, 小山内 誠, 千葉 英樹, 澤田 典均

日本癌学会総会記事 ( (一社)日本癌学会 )  64回   368 - 368  2005年09月

ヒトアデノイドにおける上皮内抗原提示細胞の分布と上皮タイト結合との関係

高野 賢一, 小島 隆, 郷 充, 新谷 朋子, 澤田 均典, 氷見 徹夫

耳鼻咽喉科免疫アレルギー ( 日本耳鼻咽喉科免疫アレルギー学会 )  23 ( 2 ) 158 - 159  2005年09月

CEAおよびDPP-IV(CD26)の局在はタイト結合蛋白occludinに依存する(Localization of carcinoembryonic antigen (CEA) and dipeptidyl peptidase IV (CD26) depend on tight junction protein occludin)

徳永 祐一, 小島 隆, 飛岡 弘敏, 村田 雅樹, 高野 賢一, 千葉 英樹, 澤田 典均

日本癌学会総会記事 ( (一社)日本癌学会 )  64回   166 - 166  2005年09月

ヒトアデノイド上皮における抗原提示細胞の分布とタイト結合の発現

小泉 純一, 小島 隆, 高野 賢一, 黒瀬 誠, 澤田 典均, 氷見 徹夫

口腔・咽頭科 ( 日本口腔・咽頭科学会 )  18 ( 1 ) 167 - 167  2005年08月

マウス骨芽細胞におけるIGF-1によるタイト結合蛋白およびギャップ結合蛋白の変化

畠山 尚子, 射場 浩介, 小島 隆, 高田 潤一, 澤田 典均, 山下 敏彦

日本整形外科学会雑誌 ( (公社)日本整形外科学会 )  79 ( 8 ) S784 - S784  2005年08月

鼻科領域の炎症研究の新しい展開 ヒト鼻粘膜におけるタイト結合による上皮バリアと抗原提示細胞

高野 賢一, 小島 隆, 澤田 典均, 氷見 徹夫

日本鼻科学会会誌 ( (一社)日本鼻科学会 )  44 ( 3 ) 185 - 185  2005年08月

マウス骨芽細胞におけるIGF-1によるタイト結合蛋白およびギャップ結合蛋白の変化

畠山 尚子, 射場 浩介, 小島 隆, 高田 潤一, 澤田 典均, 山下 敏彦

日本骨代謝学会学術集会プログラム抄録集 ( (一社)日本骨代謝学会 )  23回   270 - 270  2005年06月

Localization of apical membrane markers carcinoembryonic antigen (CEA) and dipeptidyl peptidase IV (CD26) depend on the expression of tight junction protein occludin in a polarized human intestinal epithelial cell line T84

Yuichi Tokunaga, Takashi Kojima, Hirotoshi Tobioka, Masaki Murata, Ken-ichi Takano, Hideki Chiba, Norimasa Sawada

CELL STRUCTURE AND FUNCTION ( JAPAN SOC CELL BIOLOGY )  30   88 - 88  2005年06月

研究発表ペーパー・要旨（国際会議）  

Knock down of Cx32 expression leads to alteration of claudins expression through activated MAP-kinase in primary cultures of rat hepatocytes

Takashi Kojima, Ken-ichi Takano, Masaki Murata, Masafumi Imamura, Hideki Chiba, Norimasa Sawada

CELL STRUCTURE AND FUNCTION ( JAPAN SOC CELL BIOLOGY )  30   89 - 89  2005年06月

研究発表ペーパー・要旨（国際会議）  

p38 MAP-kinase regulates function of gap and tight junctions during regeneration of rat hepatocytes

T Yamamoto, T Kojima, M Murata, K Takano, M Go, N Hatakeyama, H Chiba, N Sawada

JOURNAL OF HEPATOLOGY ( ELSEVIER SCIENCE BV )  42 ( 5 ) 707 - 718  2005年05月

　概要を見る

Background/Aims: Hepatocyte regeneration is considered to be associated with adaptive changes in expression of gap and tight junctions through multiple signal transduction pathways including p38 MAP-kinase. The role of the stress responsitive MAP-kinase, p38 MAP-kinase, signaling pathway in function of gap and tight junctions was examined during regeneration of rat hepatocytes in vivo and in vitro. Methods: We examined changes in formation, expression and function of gap and tight junctions in rat livers after 70% partial hepatectomy and in primary cultures of rat hepatocytes, by using a p38 MAP-kinase inhibitor, SB203580. Results: When p38 MAP-kinase was activated during partial hepatectomy, down-regulation of Cx32 and up-regulation of claudin-1 were observed. By SB203580 treatment, the down-regulation of Cx32 was inhibited and the up-regulation of claudin-1 was enhanced, well maintaining the structures of gap and tight junctions. SB203580 treatment did not affect the increase of hepatocyte proliferation. In EGF induced proliferative rat hepatocytes treated with SB203580, the expression and function of Cx32 and claudin-1 were increased. Conclusions: Dynamic changes of formation of gap and tight junctions during regeneration of rat hepatocytes in vivo and in vitro are in part controlled via a p38 MAP-kinase signaling pathway, and are independent of cell growth. © 2005 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.

DOI

Glyceraldehyde-derived advanced glycation end-products preferentially induce VEGF expression and reduce GDNF expression in human astrocytes

H Miyajima, M Osanai, H Chiba, N Nishikiori, T Kojima, K Ohtsuka, N Sawada

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS ( ACADEMIC PRESS INC ELSEVIER SCIENCE )  330 ( 2 ) 361 - 366  2005年05月

　概要を見る

The blood-brain barrier (BBB) is a biological unit composed of capillary endothelial cells and astrocytes. Here we examined the effects of various types of advanced glycation end-products (AGEs) on astrocytes and BBB-forming endothelial cells. While no type of AGE we examined changed the permeability of endothelial sheets, glyceraldehyde-derived AGE induced VEGF expression most significantly in astrocytes. The expression of glial cell line-derived neurotrophic factor (GDNF), which reduces the vascular permeability, was decreased in the astrocytes by treatment with glyceraldehyde-derived AGE. These results indicate that glyceraldehyde-derived AGE is the biologically active substance for astrocytes by regulating the VEGF and GDNF expression, which is causally contributing to an increase in the permeability of the BBB. (c) 2005 Elsevier Inc. All rights reserved.

DOI PubMed CiNii

HLA-DR- and CD11c-positive dendritic cells penetrate beyond well-developed epithelial tight junctions in human nasal mucosa of allergic rhinitis

K Takano, T Kojima, M Go, M Murata, S Ichimiya, T Himi, N Sawada

JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY ( HISTOCHEMICAL SOC INC )  53 ( 5 ) 611 - 619  2005年05月

　概要を見る

The epithelial barrier of the upper respiratory tract plays a crucial role in host defense. In this study, to elucidate whether there is antigen monitoring by dendritic cells (DCs) beyond the epithelial tight-junction barrier in allergic rhinitis, we investigated the expression and function of tight junctions and characterized DCs in the epithelium of nasal mucosa from patients with allergic rhinitis. In reverse transcription-polymerase chain reaction, mRNAs of tight-junction proteins occludin, JAM-1, ZO-1, and claudin-1, -4, -7, -8, -12, -13, and -14 were detected in the nasal mucosa. Occludin, JAM-1, and ZO-1 were colocalized in the uppermost layer in the pseudostratified epithelium of the nasal mucosa, whereas claudin-1, -4, and -7 were found throughout the epithelium. in freeze-fracture replicas of the nasal mucosa, continuous tight-junction strands formed well-developed networks. Epithelial barrier function measured by a dye tracer was well maintained in occludin-positive tight junctions in the epithelium of the nasal mucosa. HLA-DR- and CD11c-positive DCs expressed claudin-1 and penetrated beyond occludin in the epithelium of the nasal mucosa with, but not without, allergic rhinitis. These results indicate that DCs may easily access antigens beyond epithelial tight junctions in the human nasal mucosa of allergic rhinitis.

DOI

タイト結合蛋白occludinは生存シグナルの伝達に重要な役割を有する

村田 雅樹, 小島 隆, 高野 賢一, 畠山 尚子, 山本 敏誠, 千葉 英樹, 澤田 典均

日本病理学会会誌 ( (一社)日本病理学会 )  94 ( 1 ) 348 - 348  2005年03月

ヒトアデノイドにおける上皮内抗原提示細胞の分布と上皮タイト結合との関係

高野 賢一, 小島 隆, 郷 充, 村田 雅樹, 千葉 英樹, 氷見 徹夫, 澤田 典均

日本病理学会会誌 ( (一社)日本病理学会 )  94 ( 1 ) 376 - 376  2005年03月

骨芽細胞の分化過程におけるタイト結合蛋白の発現とその機能について

畠山 尚子, 小島 隆, 村田 雅樹, 高野 賢一, 千葉 英樹, 澤田 典均

日本病理学会会誌 ( (一社)日本病理学会 )  94 ( 1 ) 277 - 277  2005年03月

Activation of p21 (CIP1/WAF1) gene expression and inhibition of cell proliferation by overexpression of hepatocyte nuclear factor-4 alpha

H Chiba, T Itoh, S Satohisa, N Sakai, H Noguchi, M Osanai, T Kojima, N Sawada

EXPERIMENTAL CELL RESEARCH ( ELSEVIER INC )  302 ( 1 ) 11 - 21  2005年01月

　概要を見る

The F9 murine embryonal carcinoma cell line provides an attractive system for studying epithelial differentiation and antiproliferative processes. We have recently established F9 cells expressing doxycycline-inducible hepatocyte nuclear factor (HNF)-4alpha and shown that HNF-4alpha triggers the gene expression of tight-junction molecules, occludin, claudin-6, and claudin-7, as well as formation of functional tight junctions and polarized epithelial morphology (Exp. Cell Res. 286, [2003] 288). Since these events were very similar to those induced by retinoids, we investigated whether HNF-4alpha, like retinoid receptors, was involved in the control of cell proliferation. We herein show that HNF-4alpha up-regulates expression of the p21 gene, but not the p15, p16, p18, p19, or p27 gene, in a p53-independent manner, and inhibits cell growth in F9 cells. Similar results were observed in rat lung endothelial cells, in which expression of HNF-4alpha is conditionally induced by doxycycline. Furthermore, we demonstrate, by reporter assay, that HNF-4alpha significantly elevates the transcriptional activity of the p21 promoter. Since, HNF-4alpha is expressed not only in the liver but also in organs containing epithelial cells, such as kidney, intestine, pancreas, and stomach, it might also play critical roles in the regulation of epithelial morphogenesis and proliferation in these organs. (C) 2004 Elsevier Inc, All rights reserved.

DOI PubMed CiNii

Expression and function of tight junctions in the crypt epithelium of human palatine tonsils

M Go, T Kojima, K Takano, M Murata, S Ichimiya, H Tsubota, T Himi, N Sawada

JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY ( HISTOCHEMICAL SOC INC )  52 ( 12 ) 1627 - 1638  2004年12月

　概要を見る

The human palatine tonsils have surface and crypt stratified epithelium and may be initiated via the epithelium to mount immune responses to various presenting antigens. Here we investigated the expression and function of tight junctions in the epithelium of human palatine tonsils from patients with tonsillar hypertrophy or recurrent tonsillitis. Occludin, ZO-1, JAM-1, and claudin-1, -3, -4, -7, -8, and -14 mRNAs were detected in tonsillar hypertrophy. Occludin and claudin-14 were expressed in the uppermost layer of the tonsil surface epithelium, whereas ZO-1, JAM-1, and claudin-1, -4, and -7 were found throughout the epithelium. In the crypt epithelium, claudin-4 was preferentially expressed in the upper layers. In freeze-fracture replicas, short fragments of continuous tight junction strands were observed but never formed networks. In the crypt epithelium of recurrent tonsillitis, the tracer was leaked from the surface regions where occludin and claudin-4 disappeared. Occludin, ZO-1, JAM-1, and claudin-1, -3, -4, and -14, but not claudin-7, mRNAs were decreased in recurrent tonsillitis compared with those of tonsillar hypertrophy. These studies suggest unique expression of tight junctions in human palatine tonsillar epithelium, and the crypt epithelium may possess an epithelial barrier different from that of the surface epithelium.

DOI

Oncogenic Raf-1 regulates epithelial to mesenchymal transition via distinct signal transduction pathways in an immortalized mouse hepatic cell line

MD Lan, T Kojima, M Osanai, H Chiba, N Sawada

CARCINOGENESIS ( OXFORD UNIV PRESS )  25 ( 12 ) 2385 - 2395  2004年12月

　概要を見る

The epithelial to mesenchymal transition (EMT) is considered to be an important event during malignant tumor progression and metastasis. Although Raf/MEK/ERK signaling causes EMT, the mechanisms, including the signaling pathways, are as yet unclear. In the present study we have examined the effects of signal transduction pathways on oncogenic Raf-1-induced EMT, using an immortalized mouse hepatic cell line. Oncogenic Raf-1-induced EMT is characterized by down-regulation of adherens and tight junctions and the reorganization of actin. An active Raf-1 gene was introduced into a mouse hepatic cell line which was then treated with the MAP kinase inhibitor PD98059, the p38 MAP kinase inhibitor SB203580, the PI3 kinase inhibitor LY294002 or the c-Src tyrosine kinase inhibitor PP2. The expression and localization of the adherens and tight junction proteins E-cadherin, occludin, ZO-1, claudin-1 and claudin-2 were determined by western blotting, RT-PCR and immunocytochemistry. The barrier function of tight junctions was assessed by measurements of transepithelial electric resistance (TER) and permeability in terms of fluxes of [C-14]mannitol and [C-14]inulin. In Raf-1-transfected cells expression of occludin and claudin-2 was markedly down-regulated at the protein and mRNA levels and the TER value was decreased, while the permeability was increased. The distribution of ZO-1, pancadherin and F-actin was changed from linear to zipper-like structures at cell borders. In Raf-1-transfected cells treated with PD98059 and SB203580, but not LY294002, expression and localization of claudin-2, but not occludin, recovered, together with barrier function, measured as the TER value. The distributions of ZO-1, pancadherin and F-actin also recovered on treatment with PD98059 and SB203580, but not LY294002. Expression and localization of occludin recovered slightly on treatment with PP2. Thus, oncogenic Raf-1 regulates EMT via distinct MAP kinase, p38 MAP kinase and c-Src tyrosine kinase signal pathways in the mouse hepatic cell line.

DOI

IL-1 beta regulates expression of Cx32, occludin, and claudin-2 of rat hepatocytes via distinct signal transduction pathways

T Yamamoto, T Kojima, M Murata, K Takano, M Go, H Chiba, N Sawada

EXPERIMENTAL CELL RESEARCH ( ACADEMIC PRESS INC ELSEVIER SCIENCE )  299 ( 2 ) 427 - 441  2004年10月

　概要を見る

The functions of gap and tight junctions are perturbed during the acute-phase response to liver injury. To elucidate the mechanism of proinflammatory cytokine IL-1beta responsible for regulation of hepatic gap and tight junctions, we analyzed expression and function of gap and tight junctions using a rat liver injury model and primary cultures of rat hepatocyte. In rat liver lobules at 24 h after thioacetamide (TAA) treatment, where some IL-1beta-positive non-parenchymal cells existed, disappearance of connexin32-positive spots at cell borders of the hepatocytes and increases of claudin-2 and occludin immunoreactivities in bile canalicular regions were observed. In primary cultures of rat hepatocytes, IL-1beta caused the disappearance of connexin32, which was reciprocal to the induction and localization of claudin-2 to cell membranes. The downregulated connexin32 expression was inhibited by treatment with a MAP-kinase inhibitor (PD98059), whereas the upregulated claudin-2 expression was blocked by p38 MAP and PI3-kinase inhibitors (SB203580 and LY294002). The changes of connexin32 and claudin-2 may be controlled at the transcriptional level via NF-kappaB, HNF-1alpha, and CDX2. Occludin was hyperphosphorylated by IL-1beta treatment and was inhibited by treatment with a PI3-kinase inhibitor. These results demonstrate that MAP-kinase, p38 MAP-kinase, and PI3-kinase are distinctly involved in the regulation of hepatic gap and tight junctions during the acute-phase response to IL-1beta. (C) 2004 Elsevier Inc. All rights reserved.

DOI PubMed CiNii

Connexin26 によるタイト結合機能の制御 : 難聴解析モデルとしてのヒト気管支上皮細胞株の応用

郷 充, 小島 隆, 高野 賢一, 澤田 典均, 氷見 徹夫

Otology Japan   14 ( 4 )  2004年09月

CiNii

生体防御におけるヒト扁桃上皮のバリア機能の調節機構の解析

郷 充, 小島 隆, 高野 賢一, 坪田 大, 澤田 典均, 氷見 徹夫

口腔・咽頭科 = Stomato-pharyngology   17 ( 1 )  2004年08月

DOI CiNii

骨芽細胞の分化過程におけるタイト結合蛋白の発現とその機能について

畠山 尚子, 射場 浩介, 小島 隆, 高田 潤一, 澤田 典均, 山下 敏彦

日本整形外科学会雑誌 ( (公社)日本整形外科学会 )  78 ( 8 ) S878 - S878  2004年08月

ギャップ結合とタイト結合 : 形態学的関係から機能的関係へ

小島 隆, 村田 雅樹, 澤田 典均

顕微鏡 ( 公益社団法人 日本顕微鏡学会 )  39 ( 2 ) 110 - 115  2004年07月

DOI CiNii

Expression profiles and functional implications of p53-like transcription factors in thymic epithelial cell subtypes

T Kikuchi, S Ichimiya, T Kojima, L Crisa, S Koshiba, A Tonooka, N Kondo, PT van der Saag, S Yokoyama, N Sato

INTERNATIONAL IMMUNOLOGY ( OXFORD UNIV PRESS )  16 ( 6 ) 831 - 841  2004年06月

　概要を見る

In this study, we investigated the localization and functional significance of p53 tumor suppressor-like molecules, p63 and p73, in human thymic epithelial cells (TECs). Immunohistochemical studies showed particular distribution profiles of p63 and p73 in thymic epithelium, in which cortical TECs preferentially expressed p63 in their nuclei whereas subcapsular and medullary TECs expressed both p63 and p73 in their nuclei. The wide distribution of p63 in TECs was further suggested by studies using TECs of primary culture. In vitro studies using two human TEC lines demonstrated that p63 was capable of up-regulating intercellular adhesion molecule-1 (ICAM-1) and enhancing the production of IL-6 and IL-8. Moreover, in vitro studies also indicated that p73, but not p63, had the capacity to induce granulocyte macrophage colony stimulating factor (GM-CSF) and granulocyte colony stimulating factor (G-CSF) in the TEC lines. These findings suggest that p63 would regulate the cell adhesive property through ICAM-1/LFA-1 interaction and the production of IL-6 and IL-8, probably in all TEC subtypes. p73 in subcapslar and medullary TECs was suggested to play a role in the regulation of the production of GM-CSF and G-CSF, which might stimulate other stromal cells such as dendritic cells, macrophages and endothelial cells around these regions.

DOI PubMed CiNii

ヒト鼻粘膜上皮及び咽頭扁桃上皮おけるタイト結合関連蛋白の発現とバリア機能

高野 賢一, 小島 隆, 郷 充, 村田 雅樹, 千葉 英樹, 氷見 徹夫, 澤田 典均

日本病理学会会誌 ( (一社)日本病理学会 )  93 ( 1 ) 369 - 369  2004年05月

p38 MAP-kinase regulates function of gap and tight junctions during regeneration of rat hepatocytes

Takashi Kojima, Toshinobu Yamamoto, Masaki Murata, Ken-ichi Takano, Mitsuru Go, Hideki Chiba, Norimasa Sawada

CELL STRUCTURE AND FUNCTION ( JAPAN SOC CELL BIOLOGY )  29   49 - 49  2004年05月

研究発表ペーパー・要旨（国際会議）  

Is occludin transducer for survival signal?

Masaki Murata, Takashi Kojima, Toshinobu Yamamoto, Kenichi Takano, Mitsuru Go, Hideki Chiba, Norimasa Sawada

CELL STRUCTURE AND FUNCTION ( JAPAN SOC CELL BIOLOGY )  29   49 - 49  2004年05月

研究発表ペーパー・要旨（国際会議）  

The sinaling transduction pathways of TGF-beta plus EGF induced EMT in primary cultures of adult rat hepatocytes

Ken-ichi Takano, Takashi Kojima, Mitsuru Go, Masaki Murata, Hideki Chiba, Norimasa Sawada

CELL STRUCTURE AND FUNCTION ( JAPAN SOC CELL BIOLOGY )  29   48 - 48  2004年05月

研究発表ペーパー・要旨（国際会議）  

Thr(203) of claudin-1, a putative phosphorylation site for MAP kinase, is required to promote the barrier function of tight junctions

M Fujibe, H Chiba, T Kojima, T Soma, T Wada, T Yamashita, N Sawada

EXPERIMENTAL CELL RESEARCH ( ACADEMIC PRESS INC ELSEVIER SCIENCE )  295 ( 1 ) 36 - 47  2004年04月

　概要を見る

Mitogen-activated protein kinase (MAPK) modulates the barrier function of tight junctions. We identified a putative phosphorylation site for MAPK at around Thr(203) (PK-PTP) in claudin-1, and determined the biological significance of this site. To this end, using the rat lung endothelial cell line RLE, we generated cells expressing doxycycline (Dox)-inducible wild-type claudin-1 and its mutant with substitution of Thr(203) to Ala, and named them RLE:rtTA:CL1 and RLE:rtTA:CLIT203A, respectively. We herein show, by measurement of transendothelial electrical resistance and paracellular flux of mannitol and inulin, that functional tight junctions were reconstituted in both cells by Dox-induced expression of claudin-1. Interestingly, the barrier functions of tight junctions were less developed in RLE:rtTA:CL1T203A cells compared with RLE:rtTA:CLl cells. Consistently, levels of both detergent-insoluble claudin-1 protein and its threonine-phosphorylation after Dox treatment were low in RLE:rtTA:CLIT203A cells compared to RLE:rtTA:CLl cells. Furthermore, pretreatment with the MAPK inhibitor PD98059 markedly suppressed the barrier function and amount of detergent-insoluble claudin-1 in Dox-exposed RLE:rtTA:CL1 cells, whereas it marginally influenced those in RLE:rtTA:CLIT203A cells. These findings indicate that Thr(203) of claudin-1 is required to enhance the barrier function of claudin-1-based tight junctions, probably via its phosphorylation and subsequent integration into tight junctions. (C) 2004 Elsevier Inc. All rights reserved.

DOI PubMed CiNii

Inhibition of MAP kinase activity moderates changes in expression and function of Cx32 but not claudin-1 during DNA synthesis in primary cultures of rat hepatocytes

Takashi Kojima, Toshinobu Yamamoto, Mengdong Lan, Masaki Murata, Ken-Ichi Takano, Mitsuru Go, Shingo Ichimiya, Hideki Chiba, Norimasa Sawada

Medical Electron Microscopy ( Springer Japan )  37 ( 2 ) 101 - 113  2004年

　概要を見る

The signal transduction pathways and activation of the MAP kinase or PI3 kinase signaling cascade regulate a variety of cellular processes, including proliferation and differentiation in hepatocytes. To elucidate the mechanisms of signal transmission required for the regulation of gap and tight junctions during DNA synthesis in rat hepatocytes, we determined changes of expression and function of gap and tight junctions of cells grown in primary culture, using inhibitors of signaling pathways for MAP kinase (PD98059) and PI3 kinase (LY294002). During the stimulation of DNA synthesis induced by epidermal growth factor (EGF), immunoreactivity and mRNAs of gap junction protein Cx32 and of tight junction protein claudin-1 markedly decreased with reduction of gap junctional intercellular communication (GJIC) and the fence function of tight junctions. In Western blots, whole-cell lysate of claudin-1 protein decreased and phosphorylated Cx32 protein in the insoluble fraction of Triton X-100 increased during the stimulation of DNA synthesis. During reinhibition of DNA synthesis, the changes of Cx32 and claudin-1 returned to control levels, as did both functions. In treatment with the inhibitors before DNA synthesis, PD98059 inhibited the changes of expression and function of Cx32, but not claudin-1, without inhibition of cell growth, whereas LY294002 completely inhibited cell growth. These findings indicate that the PI3 kinase pathway rather than the MAP kinase pathway plays an important role for EGF-induced proliferation of rat hepatocytes, and that changes of Cx32 in hepatocytes during the stimulation of DNA synthesis may be in part controlled through MAP kinase. Furthermore, Cx32, but not claudin-1, protein may be a target of activated MAP kinase in hepatocytes.

DOI PubMed

Thr(207) of claudin-5 is involved in size-selective loosening of the endothelial barrier by cyclic AMP.

Exp Cell Res   15   201 - 12  2004年

Tight junction protein MAGI-1 is up-regulated by transfection with connexin 32 in an immortalized mouse hepatic cell line: cDNA microarray analysis.

Cell Tissue Res   19   0-0  2004年

Cyclic AMP induces phosphorylation of claudin-5 immunoprecipitates and expression of claudin-5 gene in blood-brain-barrier endothelial cells via protein kinase A-dependent and -independent pathways

T Ishizaki, H Chiba, T Kojima, M Fujibe, T Soma, H Miyajima, K Nagasawa, Wada, I, N Sawada

EXPERIMENTAL CELL RESEARCH ( ACADEMIC PRESS INC ELSEVIER SCIENCE )  290 ( 2 ) 275 - 288  2003年11月

　概要を見る

Cyclic AMP (cAMP) promotes functions of tight junctions in endothelial cells, although its target remains unknown. We showed here that cAMP increased gene expression of claudin-5 and decreased that of claudin-1 in porcine blood-brain-barrier endothelial cells via protein kinase A (PKA)-independent and -dependent pathways, respectively. cAMP also enhanced immunoreactivity of claudin-5 along cell borders and in the cytoplasm, reorganized actin filaments, and altered signals of claudin-5, occludin, ZO-1, and ZO-2 along cell boundaries from zipperlike to linear patterns. In contrast, claudin-1 was detected only in the cytoplasm in a dotlike pattern, and its immunolabeling was reduced by cAMP. Interestingly, 31- and 62-kDa claudin-5 immunoprecipitates in the NP-40-soluble and -insoluble fractions, respectively, were highly phosphorylated on threonine residue(s) upon cAMP treatment. All these changes induced by cAMP, except for claudin-5 expression and its signals in the cytoplasm, were reversed by an inhibitor of PKA, H-89. We also demonstrated that cAMP elevated the barrier function of tight junctions in porcine blood-brain-barrier endothelial cells in PKA-dependent and -independent manners. These findings indicate that both PKA-induced phosphorylation of claudin-5 immunoprecipitates and cAMP-dependent but PKA-independent induction of claudin-5 expression could be involved in promotion of tight-junction function in endothelial cells. (C) 2003 Elsevier Inc. All rights reserved.

DOI PubMed CiNii

Regulation of the blood-biliary barrier: Interaction between gap and tight junctions in hepatocytes

Takashi Kojima, Toshinobu Yamamoto, Masaki Murata, Hideki Chiba, Yasuo Kokai, Norimasa Sawada

Medical Electron Microscopy   36 ( 3 ) 157 - 164  2003年09月

書評論文，書評，文献紹介等  

　概要を見る

Hepatocytes tightly connect with each other by intercellular junctions to form liver cell plates. The junctions composed of gap, tight, and adherens junctions and desmosomes concentrate around bile canaliculi. In particular, tight junctions serve as a barrier to keep bile in bile canaliculi away from the blood circulation. Thus, it is very reasonable to call tight junctions of hepatocytes the blood-biliary barrier. On the other hand, gap junctions of hepatocytes are considered to enable ordered contraction of bile canaculi from centrizonal to periportal hepatocytes by their function of intercellular communication. Gap and tight junctions may thus play a crucial role in bile secretion, one of the most differentiated functions of the liver. In intrahepatic cholestasis, a common pathological condition of the liver, downregulation of gap and tight junctional functions is seen, which results in impaired intercellular communication and in leaky tight junctions. Although the changes in gap and tight junctions had been considered to be independent of each other, recent findings that the tight junction-associated proteins ZO-1 and occludin bind to connexins indicate the possibility of either coordinate or reciprocal regulation of macromolecular complexes containing gap- and tight-junction proteins. In this review, we introduce the interaction and regulation between gap and tight junctions of hepatocytes in vitro and discuss the regulatory mechanisms of the "blood-biliary barrier" to study the molecular pathogenesis of cholestasis.

DOI PubMed CiNii

Tight junctions and human diseases

Norimasa Sawada, Masaki Murata, Keisuke Kikuchi, Makoto Osanai, Hirotoshi Tobioka, Takashi Kojima, Hideki Chiba

Medical Electron Microscopy   36 ( 3 ) 147 - 156  2003年09月

書評論文，書評，文献紹介等  

　概要を見る

Tight junctions are intercellular junctions adjacent to the apical end of the lateral membrane surface. They have two functions, the barrier (or gate) function and the fence function. The barrier function of tight junctions regulates the passage of ions, water, and various macromolecules, even of cancer cells, through paracellular spaces. The barrier function is thus relevant to edema, jaundice, diarrhea, and blood-borne metastasis. On the other hand, the fence function maintains cell polarity. In other words, tight junctions work as a fence to prevent intermixing of molecules in the apical membrane with those in the lateral membrane. This function is deeply involved in cancer cell biology, in terms of loss of cell polarity. Of the proteins comprising tight junctions, integral membrane proteins occludin, claudins, and JAMs have been recently discovered. Of these molecules, claudins are exclusively responsible for the formation of tight-junction strands and are connected with the actin cytoskeleton mediated by ZO-1. Thus, both functions of tight junctions are dependent on the integrity of the actin cytoskeleton as well as ATP. Mutations in the claudin14 and the claudin16 genes result in hereditary deafness and hereditary hypomagnesemia, respectively. Some pathogenic bacteria and viruses target and affect the tight-junction function, leading to diseases. In this review, the relationship between tight junctions and human diseases is summarized.

DOI PubMed CiNii

Role of the p38 MAP-kinase signaling pathway for Cx32 and claudin-1 in the rat liver

T Kojima, T Yamamoto, M Murata, M Lan, KI Takano, M Go, S Ichimiya, H Chiba, N Sawada

CELL COMMUNICATION AND ADHESION ( TAYLOR & FRANCIS LTD )  10 ( 4-6 ) 437 - 443  2003年07月

　概要を見る

Liver regeneration and cholestasis are associated with adaptive changes in expression of gap and tight junctions through signal transduction. The roles of stress responsitive MAP-kinase, p38 MAP-kinase, in the signaling pathway for gap junction protein, Cx32, and tight junction protein, claudin-1, were examined in rat liver in vivo and in vitro , including regeneration following partial hepatectomy and cholestasis after common bile duct ligation. Changes in the expression and function of Cx32 and claudin-1 in hepatocytes in vivo were studied using the p38 MAP-kinase inhibitor SB203580. Following partial hepatectomy and common bile duct ligation, down-regulation of Cx32 protein was inhibited by SB203580 treatment. Up-regulation of claudin-1 protein was enhanced by SB203580 treatment after partial hepatectomy but not common bile duct ligation. However, no change of the Ki-67 labeling index (which is a marker for cell proliferation) in the livers treated with SB203580, was observed compared to that without SB203580 treatment. In primary cultures of rat hepatocytes, however, treatment with a p38 MAP-kinase activator, anisomycin, decreased Cx32 and claudin-1 protein levels. p38 MAP-kinase may be an important signaling pathway for regulation of gap and tight junctions in hepatocytes. Changes of gap and tight junctions during liver regeneration and cholestasis are shown to be in part controlled via the p38 MAP-kinase signaling pathway and are independent of cell growth.

DOI

Hepatocyte nuclear factor (HNF)-4 alpha triggers formation of functional tight junctions and establishment of polarized epithelial morphology in F9 embryonal carcinoma cells

H Chiba, T Gotoh, T Kojima, S Satohisa, K Kikuchi, M Osanai, N Sawada

EXPERIMENTAL CELL RESEARCH ( ACADEMIC PRESS INC ELSEVIER SCIENCE )  286 ( 2 ) 288 - 297  2003年06月

　概要を見る

F9 murine embryonal carcinoma cells provide an attractive system for facilitating molecular mechanisms for epithelial morphogenesis, since they have the capability of differentiating into polarized epithelial cells bearing an apical junctional complexes. We previously showed that a specific retinoid X receptor-retinoic acid receptor heterodimer transduced retinoid signals for biogenesis of functional tight junctions in F9 cells (Exp. Cell Res. 263, (2001) 163). In the present study we generated F9 cells expressing doxycycline-inducible hepatocyte nuclear factor (HNF)-4alpha, a nuclear receptor. We herein show that induction of HNF-4alpha initiates differentiation of F9 cells to polarized epithelial cells, in which tight-junction proteins occludin, claudin-6, claudin-7, and ZO-1 are concentrated at the apical-most regions of lateral membranes. Expression of occludin, claudin-6, and claudin-7 was induced in the cells by doxycycline treatment in a dose- and time-dependent manner, in terms of the amount of HNF-4a. In contrast, expression levels of ZO-1, ZO-2, E-cadherin, and P-catenin were not altered by HNF-4a. We also demonstrate, by analysis of diffusion of labeled sphingomyelin, that the fence function of tight junctions is achieved by induction of HNF-4a. These findings indicate that HNF-4a triggers de novo formation of functional tight junctions and establishment of epithelial cell polarity. (C) 2003 Elsevier Science (USA). All rights reserved.

DOI PubMed CiNii

細胞間の接着とシグナル Hepatocyte nuclear factor(HNF)-4αはタイト結合分子の発現を制御し,上皮の分化と極性形成を誘導する

千葉 英樹, 郷久 晴朗, 小島 隆, 菊地 慶介, 小山内 誠, 澤田 典均

日本細胞生物学会大会講演要旨集 ( (一社)日本細胞生物学会 )  56回   18 - 18  2003年05月

高カリウム培地とouabainによるヒト気管支上皮細胞(Calu-3)のタイト結合の変化

郷 充, 小島 隆, 村田 雅樹, 高野 賢一, 山本 敏誠, 千葉 英樹, 澤田 典均

日本細胞生物学会大会講演要旨集 ( (一社)日本細胞生物学会 )  56回   34 - 34  2003年05月

IL-1βによる急性期反応時の肝ギャップ及びタイト結合の変化はMAP-,p38MAP-及びPI3-kinase経路を介して制御されている

山本 敏誠, 小島 隆, 高野 賢一, 村田 雅樹, 郷 充, 千葉 英樹, 小海 康夫, 澤田 典均

日本細胞生物学会大会講演要旨集 ( (一社)日本細胞生物学会 )  56回   32 - 32  2003年05月

肝細胞のタイト結合における膜貫通タイト結合蛋白occludin分子の役割

村田 雅樹, 小島 隆, 山本 敏誠, 高野 賢一, 郷 充, 千葉 英樹, 小海 康夫, 澤田 典均

日本細胞生物学会大会講演要旨集 ( (一社)日本細胞生物学会 )  56回   33 - 33  2003年05月

肝細胞におけるコネキシン32によるタイト結合誘導のメカニズム

小島 隆, 村田 雅樹, 山本 敏誠, 郷 充, 高野 賢一, 千葉 英樹, 澤田 典均

日本細胞生物学会大会講演要旨集 ( (一社)日本細胞生物学会 )  56回   33 - 33  2003年05月

肝細胞のタイト結合におけるoccludinの役割

村田 雅樹, 小島 隆, 山本 敏誠, 千葉 英樹, 小海 康夫, 澤田 典均

日本病理学会会誌 ( (一社)日本病理学会 )  92 ( 1 ) 335 - 335  2003年04月

ストレス応答MAPキナーゼ経路を介した肝細胞のギャップ及びタイト結合の発現調節機構

小島 隆, 山本 敏誠, 村田 雅樹, 神原 由季, 小海 康夫, 千葉 英樹, 澤田 典均

日本病理学会会誌 ( (一社)日本病理学会 )  92 ( 1 ) 336 - 336  2003年04月

"すきま"の細胞生物学 細胞間接着装置タイト結合とヒト疾患

澤田 典均, 小島 隆, 飛岡 弘敏, 小海 康夫, 千葉 英樹

札幌医学雑誌 ( 札幌医科大学 )  72 ( 1～2 ) 1 - 7  2003年04月

Epigenetic inactivation of CHFR and sensitivity to microtubule inhibitors in gastric cancer.

Cancer Res   63   8606 - 13  2003年

TGF-beta/Smadsシグナルによる肝細胞のギャップ及びタイト結合の変化

小島 隆, 山本 敏誠, 小海 康夫, 千葉 英樹, 澤田 典均

日本癌学会総会記事 ( (一社)日本癌学会 )  61回   135 - 135  2002年10月

IL-1βは肝細胞のギャップ及びタイト結合の発現を相反的に調節している

山本 敏誠, 小島 隆, 小海 康夫, 千葉 英樹, 澤田 典均

日本癌学会総会記事 ( (一社)日本癌学会 )  61回   134 - 134  2002年10月

Cx32 formation and/or Cx32-mediated intercellular communication induces expression and function of tight junctions in hepatocytic cell line

T Kojima, DC Spray, Y Kokai, H Chiba, Y Mochizuki, N Sawada

EXPERIMENTAL CELL RESEARCH ( ACADEMIC PRESS INC ELSEVIER SCIENCE )  276 ( 1 ) 40 - 51  2002年05月

　概要を見る

Gap junctional intercellular communication (GJIC) is thought to play a crucial role in cell differentiation. Small gap junction plaques are frequently associated with tight junction strands in hepatocytes, suggesting that gap junctions may be closely related to the role of tight junctions in the establishment of cell polarity. To examine the exact role of gap junctions in regulating tight junctions, we transfected connexin 32 (Cx32), Cx26, or Cx43 cDNAs into immortalized mouse hepatocytes derived from Cx32-deficient mice and examined the expression and function of the endogenous tight junction molecules. In transient wild-type Cx32 transfectants, immunocytochemistry revealed that endogenous occludin was in part localized at cell borders, where it was colocalized with Cx32, whereas neither was detected in parental cells. In Cx32 null hepatocytes transfected with Cx32 truncated at position 220 (R220stop), wild-type Cx26, or wild-type Cx43 cl)NAs, occludin was not detected at cell borders. In stable wild-type Cx32 transfectants, occludin, claudin-1, and ZO-1 mRNAs and proteins were significantly increased compared to parental cells and all of the proteins were colocalized with Cx32 at cell borders. Treatment with a GJIC blocker, 18beta-glycyrrhetinic acid, resulted in decreases of occludin and claudin-1 at cell borders in the stable transfectants. The induction of tight junction proteins in the stable transfectants was accompanied by an increase in both fence and barrier functions of tight junctions. Furthermore, in the stable transfectants, circumferencial actin filaments were also increased without a change of actin protein. These results indicate that Cx32 formation and/or Cx32-mediated intercellular communication may participate in the formation of functional tight junctions and actin organization. (C) 2002 Elsevier Science (USA).

DOI PubMed CiNii

Hepatocyte nuclear factor (HNF)-4 alpha induces expression of endothelial Fas ligand (FasL) to prevent cancer cell transmigration: A novel defense mechanism of endothelium against cancer metastasis

M Osanai, H Chiba, T Kojima, M Fujibe, K Kuwahara, H Kimura, M Satoh, N Sawada

JAPANESE JOURNAL OF CANCER RESEARCH ( BUSINESS CENTER ACADEMIC SOCIETIES JAPAN )  93 ( 5 ) 532 - 541  2002年05月

　概要を見る

Endothelial Fas ligand (FasL) contributes to the "immune privilege" of tissues such as testis and eye, in which apoptosis is induced in infiltrating Fas-positive activated T cells and results in the inhibition of leukocyte extravasation. In this study, we examined the role of endothelial FasL in controlling cancer cell transmigration using rat lung endothelial (RLE) cell line bearing a doxycycline-inducible hepatocyte nuclear factor (HNF)-4alpha expression system. We showed that a detectable level of FasL was expressed in RLE cells and that this expression was markedly up-regulated and well correlated to the degree of HNF-4alpha expression in a time-dependent manner. When various cancer cells were overlaid on an RLE monolayer sheet, we examined the ability of endothelial FasL to induce massive apoptosis in Fas-expressing cancer cells and found a causal link to inhibition of the transmigration. Finally we showed that FasL was expressed in capillaries of the rat brain by immunohistochemical staining, suggesting that FasL serves its functions not only in vitro, but also in vivo. These results raise the possibility that HNF-4alpha is involved in regulating cancer cell transmigration by modulating the Fas-FasL system.

p73 is expressed in human thymic epithelial cells

S Ichimiya, T Kojima, H Momota, N Kondo, T Ozaki, A Nakagawara, ML Toribio, M Imamura, N Sato

JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY ( HISTOCHEMICAL SOC INC )  50 ( 4 ) 455 - 462  2002年04月

　概要を見る

The thymus is a heterogeneous immune organ in which immature T-cells develop and eventually specialize to make certain immune responses of their own. Among various types of stromal cells in the thymus, thymic epithelial cells (TECs) have a crucially important function for presenting self-antigens and secreting cytokines to thymocytes for their maturation into T-cells. In this study we show that the p73 gene, a homologue of the tumor suppressor gene p53, was expressed in the nucleus of the human TEC in vivo and in TEC lines in vitro. Because p73 has the capacity to be a transactivator like p53, it may contribute to T-cell development in the context of TEC biology as regulated in the cell cycle and apoptosis.

The p53 family member genes are involved in the notch signal pathway

Y Sasaki, S Ishida, Morimoto, I, T Yamashita, T Kojima, C Kihara, T Tanaka, K Imai, Y Nakamura, T Tokino

JOURNAL OF BIOLOGICAL CHEMISTRY ( AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC )  277 ( 1 ) 719 - 724  2002年01月

　概要を見る

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.

DOI PubMed CiNii

Growth-suppressive function of human connexin32 in a conditional immortalized mouse hepatocyte cell line

T Kojima, M Srinivas, A Fort, M Urban, GH Lee, N Sawada, DC Spray

IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL ( SOC IN VITRO BIOLOGY )  37 ( 9 ) 589 - 598  2001年10月

　概要を見る

Mouse hepatocytes immortalized with a temperature-sensitive allele of the SV40 large T-antigen (CHST8 cells) were found to lack the high expression of the gap junction proteins Gx26 and Cx32 that characterizes normal mouse hepatocytes. expressing instead Cx43 and Cx45 at minimal levels. In order to examine the growth suppressive function of Cx32 on hepatocytes we transfected these CHST8 cells with human Cx32 complementary deoxyribonucleic acid and measured the growth rates at 33, 37, and 39 degrees C. Expression of human Cx32 and its messenger ribonucleic acid in the stable cell lines was confirmed by immunocytochemistry and by Western and Northern blots analyses. Dye transfer following lucifer yellow injection into the transfectants was extensive; Cx32 channels displayed unitary conductances of about 70 pS and were moderately voltage sensitive. When cultured at 33 and 39 degrees C, growth rates of both parental cells and transfectants were of the same level. When examined at 37 degrees C. growth rate of the transfectant, which highly expressed Cx32 at the membranes, was significantly decreased compared to the parental cells. However. no changes in the expression of Cx32 protein in the transfectants were observed between 33 and 37 degrees C. These results suggest that Cx32 expression could inhibit hepatocyte growth in vitro using the conditional immortalized cells. Cx32 transfectants using a conditional immortalized mouse hepatocyte may be useful for examining the mechanisms of growth and differentiation in hepatocytes by gap junction expression.

DOI

Global ischemia-induced increases in the gap junctional proteins connexin 32 (Cx32) and Cx36 in hippocampus and enhanced vulnerability of Cx32 knock-out mice

K Oguro, T Jover, H Tanaka, Y Lin, T Kojima, N Oguro, SY Grooms, MVL Bennett, RS Zukin

JOURNAL OF NEUROSCIENCE ( SOC NEUROSCIENCE )  21 ( 19 ) 7534 - 7542  2001年10月

　概要を見る

Gap junctions are conductive channels that connect the interiors of coupled cells. In the hippocampus, GABA-containing hippocampal interneurons are interconnected by gap junctions, which mediate electrical coupling and synchronous firing and thereby promote inhibitory transmission. The present study was undertaken to examine the hypothesis that the gap junctional proteins connexin 32 (Cx32; expressed by oligodendrocytes, interneurons, or both), Cx36 (expressed by interneurons), and Cx43 (expressed by astrocytes) play a role in defining cell-specific patterns of neuronal death in hippocampus after global ischemia in mice. Global ischemia did not significantly alter Cx32 and Cx36 mRNA expression and slightly increased Cx43 mRNA expression in the vulnerable CA1, as assessed by Northern blot analysis and in situ hybridization. Global ischemia induced a selective increase in Cx32 and Cx36 but not Cx43 protein abundance in CA1 before onset of neuronal death, as assessed by Western blot analysis. The increase in Cx32 and Cx36 expression was intense and specific to parvalbumin-positive inhibitory interneurons of CA1, as assessed by double immunofluorescence. Protein abundance was unchanged in CA3 and dentate gyrus. The finding of increase in connexin protein without increase in mRNA suggests regulation of Cx32 and Cx36 expression at the translational or post-translational level. Cx32(Y/-) null mice exhibited enhanced vulnerability to brief ischemic insults, consistent with a role for Cx32 gap junctions in neuronal survival. These findings suggest that Cx32 and Cx36 gap junctions may contribute to the survival and resistance of GABAergic interneurons, thereby defining cell-specific patterns of global ischemia-induced neuronal death.

Gap junction expression and cell proliferation in differentiating cultures of Cx43 KO mouse hepatocytes

T Kojima, A Fort, MY Tao, M Yamamoto, DC Spray

AMERICAN JOURNAL OF PHYSIOLOGY-GASTROINTESTINAL AND LIVER PHYSIOLOGY ( AMER PHYSIOLOGICAL SOC )  281 ( 4 ) G1004 - G1013  2001年10月

　概要を見る

Primary cultures of adult mouse hepatocytes are shown here to reexpress differentiated hepatocyte features following treatment with 2% DMSO and 10(-7) M glucagon. To examine the roles of gap junctional communication during hepatocyte growth and differentiation, we have compared treated and untreated hepatocytes from connexin (Cx) 32-deficient [Cx32 knockout (KO)] and wild-type mice. In untreated cultures, DNA replication of Cx32 KO hepatocytes was markedly higher than of wild types. Although Cx26 mRNA levels remained high at all time points in wild-type and Cx32 KO hepatocytes, Cx32 mRNA and protein in wild-type hepatocytes underwent a marked decline, which recovered in 10-day treated cultures. Increased levels of Cx26 protein and junctional conductance were observed in Cx32 KO hepatocytes at 96 h in culture, a time when cell growth rate was high. Treatment with DMSO/glucagon highly reinduced Cx26 expression in Cx32 KO hepatocytes, and such treatment reinduced expression of both Cx32 and Cx26 expression in wild types. Dye transfer was not observed following Lucifer yellow injection into DMSO/glucagon-treated Cx32 KO hepatocytes, whereas the spread was extensive in wild types. Nevertheless, high junctional conductance values were observed in treated cells from both genotypes. These studies provide a method by which the differentiated phenotype can be obtained in cultured mouse hepatocytes and provide in vitro evidence that expression of gap junctions formed of Cx32 are involved in the regulation of growth of mouse hepatocytes.

肝細胞のギャップ及びタイト結合の発現調節におけるp38MAP-kinase pathwayの関与

小島 隆, 小海 康夫, 千葉 英樹, 澤田 典均

日本癌学会総会記事 ( (一社)日本癌学会 )  60回   93 - 93  2001年09月

初代培養ラット肝細胞のギャップ及びタイト結合の発現調節に対するMAP kinase及びIP3 kinaseの抑制物質の影響

小島 隆, 小海 康夫, 千葉 英樹, 桑原 和英, 澤田 典均

日本細胞生物学会大会講演要旨集 ( (一社)日本細胞生物学会 )  54回   94 - 94  2001年05月

Occludin and claudin-1 concentrate in the midbody of immortalized mouse hepatocytes during cell division

T Kojima, Y Kokai, H Chiba, M Osanai, K Kuwahara, M Mori, Y Mochizuki, N Sawada

JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY ( HISTOCHEMICAL SOC INC )  49 ( 3 ) 333 - 339  2001年03月

　概要を見る

It has been believed that epithelial cells maintain tight junctions at all times, including during cell division, to provide a continuous epithelial seal. However, changes in localization of integral tight junction proteins during cell division have not been examined. In this study, using SV40-immortalized mouse hepatocytes transfected with human Cx32 cDNA, in which tight junction strands and the endogenous tight junction proteins occludin, claudin-1, ZO-1, and ZO-2 were induced, we examined changes in localization of the tight junction proteins at all stages of cell division. All tight junction proteins were present between mitotic cells and neighboring cells throughout cell division. in late telophase, the integral tight junction proteins occludin and claudin-1, but not the cytoplasmic proteins ZO-1 and ZO-2, were concentrated in the midbody between the daughter cells and were observed at cell borders between the daugher and neighboring cells. These results indicate that the integral tight junction proteins are regulated in a different manner from the cytoplasmic proteins ZO-1 and ZO-2 during cytokinesis.

Cx32 but not Cx26 is associated with tight junctions in primary cultures of rat hepatocytes

T Kojima, Y Kokai, H Chiba, M Yamamoto, Y Mochizuki, N Sawada

EXPERIMENTAL CELL RESEARCH ( ACADEMIC PRESS INC )  263 ( 2 ) 193 - 201  2001年02月

　概要を見る

On freeze-fracture replicas, gap junctions are frequently colocalized with tight junctions. In this study, to elucidate the relationship between gap- and tight-junction proteins, we investigated the localization of gap-junction proteins Cx32 and Cx26 and tight-junction proteins occludin, claudin-1, ZO-1, and ZO-2 in primary cultured rat hepatocytes, using confocal laser microscopy. In hepatocytes cultured in 2% DMSO and 10(-7) M glucagon medium, Cx32- but not Cx26-immunoreactive lines were observed on the most sukapical plasma membrane at cell borders, while on the basolateral membrane both Cx32- and Cx26-positive spots were colocalized. Occludin-, claudin-1-, ZO-1-, and ZO-2-immunoreactive lines were also linearly observed on the most subapical plasma membrane and were colocalized with only Cx32-immunoreactive lines. In freeze-fracture analysis, many small gap-junction plaques were observed within a well-developed tight-junction strand network. The fence function of tight junctions in the cells, as examined by diffusion of labeled sphingomyelin, was well maintained. We also carried out Western blotting for Cx32 following immunoprecipitation with anti-occludin, anti-claudin-1, or anti-ZO-1 antibodies. Cx32 was detectable in all immunoprecipitates. These results suggest that Cx32 gap junctions, but not those with Cx26, are closely coordinated with the expression and function of tight junctions in hepatocytes and that Cx32 gap-junction formation may affect cell polarity through modification of tight-junction expression. (C) 2001 Academic Press.

DOI PubMed CiNii

[Development of hepatocellular carcinoma during chronic hepatitis]

Nippon Rinsho.   59   89 - 94  2001年

Occludin regulates actin cytoskeleton in endothelial cells.

Cell Struct Funct   26 ( 2 ) 109 - 116  2001年

DOI PubMed CiNii

Gap and tight junctions in liver: composition, regulation and function

In The Liver: Biology and Pathobiology, 4th ed    2001年

A close relation of occludin to actin cytoskeleton in endothelial cells

Cell struct. Funct   26   101 - 108  2001年

慢性肝炎から肝癌へ：その分子生物学機序

日本臨床   59   89 - 94  2001年

ギャップ結合と細胞分化：ギャップ結合蛋白（Cx32）によるタイト結合の誘導およびアクチン構築の変化

肝サイトスケルトン研究会誌   11   17 - 22  2001年

Cx32 formation and/or Cx32 mediated intercellular communication induce tight junctions in hepatocytes

T Kojima, Y Kokai, H Chiba, M Yamamoto, D Spray, Y Mochizuki, N Sawada

MOLECULAR BIOLOGY OF THE CELL ( AMER SOC CELL BIOLOGY )  11   330A - 330A  2000年12月

研究発表ペーパー・要旨（国際会議）  

ギャップ結合と細胞分化 ギャップ結合によるタイト結合の誘導

小島 隆, 千葉 英樹, 小海 康夫, 望月 洋一, 澤田 典均

日本癌学会総会記事 ( (一社)日本癌学会 )  59回   36 - 36  2000年09月

ギャップ結合とタイト結合の形成過程における相互関係

山本 正夫, 小島 隆

日本臨床電子顕微鏡学会誌 ( 日本臨床分子形態学会 )  33 ( 増刊 ) S81 - S81  2000年08月

培養肝細胞におけるギャップ結合蛋白(C×32)によるタイト結合の誘導

小島 隆, 望月 洋一, 澤田 典均, 小海 康夫, 千葉 英樹

日本病理学会会誌 ( (一社)日本病理学会 )  89 ( 1 ) 174 - 174  2000年03月

Uncoupling of gate and fence functions of MDCK cells by the actin depolymerizing reagent mycalolide B

Exp. Cell Res   257 ( 2 ) 238 - 244  2000年

DOI PubMed CiNii

"Negative" physiology: What connexin-deficient mice reveal about the functional roles of individual gap junction proteins

DC Spray, T Kojima, E Scemes, SO Suadicani, Y Gao, S Zhao, A Fort

GAP JUNCTIONS ( ELSEVIER ACADEMIC PRESS INC )  49   509 - 533  2000年

書評論文，書評，文献紹介等  

DOI

Induction of tight junctions in human connexin 32 (hCx32)-transfected mouse hepatocytes: Connexin 32 interacts with occludin

T Kojima, N Sawada, H Chiba, Y Kokai, M Yamamoto, M Urban, GH Lee, EL Hertzberg, Y Mochizuki, DC Spray

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS ( ACADEMIC PRESS INC )  266 ( 1 ) 222 - 229  1999年12月

　概要を見る

Small gap junction plaques are associated with tight junction strands in some cell types including hepatocytes and it is thought that they may be closely related to tight junctions and the establishment of cell polarity. In order to examine roles of gap junctions in regulating expression and structure of tight junctions, we transfected human Cx32 cDNA into immortalized mouse hepatocytes (CHST8 cells) which lack endogenous Cx32 and Cx26. Immunocytochemistry revealed that endogenous integral tight junction protein occludin was strongly localized and was colocalized with Cx32 at cell borders in transfectants, whereas neither was detected in parental cells. In Northern blots, mRNAs encoding occludin and the other integral tight junction proteins, claudin-1 and -2, were induced in the transfectants compared to parental cells. In Western blots, occludin protein was increased in the transfectants compared to parental cells, and binding of occludin to Cx32 protein was demonstrated by immunoprecipitation. In freeze fracture of the transfectants, tight junction strands were more numerous and complex compared to parental cells, and small gap junction plaques appeared within induced tight junction strands. Nevertheless, no change in barrier function of tight junctions was observed. These results indicate that in hepatocytes, gap junction, and tight junction expression are closely coordinated, and that Cx32 may play a role in regulating occludin expression. (C) 1999 Academic Press.

DOI PubMed CiNii

Functional properties of channels formed by the neuronal gap junction protein connexin36

M Srinivas, R Rozental, T Kojima, R Dermietzel, M Mehler, DF Condorelli, JA Kessler, DC Spray

JOURNAL OF NEUROSCIENCE ( SOC NEUROSCIENCE )  19 ( 22 ) 9848 - 9855  1999年11月

　概要を見る

The expression and functional properties of connexin36 (Cx36) were examined in two communication-deficient cell lines (N2A-neuroblastoma and PC-12 cells) transfected with Cx36 and in hippocampal neurons that express the connexin endogenously. Transfected cells expressed the expected 2.9 kb Cx36 transcript and Cx36 immunoreactivity, whereas nontransfected cells were devoid of Cx36. The relationship between steady-state junctional conductance (g(j)) and transjunctional voltage was well described by a two-state Boltzmann equation. The half-inactivation voltage (V-0), the ratio of minimal to maximal g(j) (g(min)/g(max)), and the equivalent gating charge were +/- 75 mV, 0.55, and 1.75, respectively, indicating that Cx36 exhibits very low voltage sensitivity. Conductance of single Cx36 channels measured with patch pipettes containing 130 mM CsCl was 10-15 pS (n = 15 cell pairs); despite this low unitary conductance, Cx36 channels were permeable to the dye Lucifer yellow. Hippocampal neurons expressed Cx36 both in vivo and in culture. The electrophysiological properties of channels in cultured hippocampal neurons were similar to those of the channels expressed by the transfected cell lines, and the neuronal channels were similarly permeable to Lucifer yellow. The unique combination of weak voltage sensitivity, small unitary conductance, and permeation by anions as large as second messenger molecules endows Cx36 gap junction channels with properties well suited for mediating flexible electrical and biochemical interactions between neurons.

Knockdown of AMPA receptor GluR2 expression causes delayed neurodegeneration and increases damage by sublethal ischemia in hippocampal CA1 and CA3 neurons

K Oguro, N Oguro, T Kojima, SY Grooms, A Calderone, Zheng, X, MVL Bennett, RS Zukin

JOURNAL OF NEUROSCIENCE ( SOC NEUROSCIENCE )  19 ( 21 ) 9218 - 9227  1999年11月

　概要を見る

Considerable evidence suggests that Ca2+-permeable AMPA receptors are critical mediators of the delayed, selective neuronal death associated with transient global ischemia and sustained seizures. Global ischemia suppresses mRNA and protein expression of the glutamate receptor subunit GluR2 and increases AMPA receptor-mediated Ca2+ influx into vulnerable neurons of the hippocampal CA1 before the onset of neurodegeneration. Status epilepticus suppresses GluR2 mRNA and protein in CA3 before neurodegeneration in this region. To examine whether acute downregulation of the GluR2 subunit, even in the absence of a neurological insult, can cause neuronal cell death, we performed GluR2 "knockdown" experiments. Intracerebral injection of antisense oligodeoxynucleotides targeted to GluR2 mRNA induced delayed death of pyramidal neurons in CA1 and CA3. Antisense-induced neurodegeneration was preceded by a reduction in GluR2 mRNA, as indicated by in situ hybridization, and in GluR2 protein, as indicated by Western blot analysis. GluR2 antisense suppressed GluR2 mRNA in the dentate gyrus but did not cause cell death. The AMPA receptor antagonist 6-cyano-7-nitroquinoxiline-2,3-dione (CNQX) and the Ca2+-permeable AMPA receptor channel blocker 1-naphthyl acetyl spermine protected against antisense-induced cell death. This result indicates that antisense-induced cell death is mediated by Ca2+-permeable AMPA receptors. GluR2 antisense and brief sublethal global ischemia acted synergistically to cause degeneration of pyramidal neurons, consistent with action by a common mechanism. These findings demonstrate that downregulation of GluR2 is sufficient to induce delayed death of specific neuronal populations.

肝ギャップ結合蛋白(Cx32)によるタイト結合の誘導

小島 隆, 澤田 典均, 森 道夫, 望月 洋一

日本臨床電子顕微鏡学会誌 ( 日本臨床分子形態学会 )  32 ( 増刊 ) S110 - S110  1999年10月

Disruption of circumferential actin filament causes disappearance of occludin from the cell borders of rat hepatocytes in primary culture without distinct changes of tight junction strands

T Kojima, N Sawada, M Yamamoto, Y Kokai, M Mori, Y Mochizuki

CELL STRUCTURE AND FUNCTION ( JAPAN SOC CELL BIOLOGY )  24 ( 1 ) 11 - 17  1999年02月

　概要を見る

We investigated the relationship of actin filament organization to occludin and tight junction strands in primary cultured rat hepatocytes using an actin depolymerizing agent, mycalolide B. In control cultures, well-developed circumferential actin filaments and occludin immunoreactivity were observed on the most subapical plasma membrane of the cells, and tight junction strands formed well-developed networks in freeze-fracture replicas. In hepatocytes treated with 3 mu M mycalolide B for 6 h, circumferential actin filaments and occludin immunoreactivity disappeared from the cell borders, However, there were no marked abnormalities of tight junction strands in freeze fracture replicas. Similar results were obtained from cells cultured in medium with 0.05 mM Ca2+ for 6 h. The close association of occludin with actin and the existence of intact tight junction strands that are virtually free of both occludin and actin suggest a physiological role of occludin, but not the other proteins forming the tight junction strands, in the linkage between actin cytoskeleton and tight junction.

DOI PubMed CiNii

Spontaneous appearance of circular actin bands in cultured hepatocytes of adult rats

MINAMITSUJI-MOCHIZUKI Chihiro, MITAKA Toshihiro, KOJIMA Takashi, NIITSU Yoshiro, MOCHIZUKI Yohichi

Medical electron microscopy : official journal of the Clinical Electron Microscopy Society of Japan   32 ( 2 ) 114 - 121  1999年

DOI PubMed CiNii

細胞接着装置 タイト結合とギャップ結合の機能測定法

新培養細胞実験法    1999年

Expression and function of the human liver gap junction protein, Cx26 in transfected cells.

T Kojima, M Srinivas, A Fort, G Lee, DC Spray

HEPATOLOGY ( W B SAUNDERS CO )  28 ( 4 ) 594A - 594A  1998年10月

研究発表ペーパー・要旨（国際会議）  

Gap junction expression and function in primary cultures of CX32 deficient (KO) mouse hepatocytes.

T Kojima, M Tao, A Fort, Z Huang, DC Spray

HEPATOLOGY ( W B SAUNDERS CO )  28 ( 4 ) 176A - 176A  1998年10月

研究発表ペーパー・要旨（国際会議）  

Occludin expression and tight junction strand formation during replicative DNA synthesis in primary cultures of rat hepatocytes

KOJIMA T

Med Electron Microsc   31 ( 4 ) 169 - 176  1998年

DOI CiNii

Effects of the protein phosphatase inhibitors, okadaic acid and vanadate, on localization of occuludin in primary cultures of rat hepatocytes

Kojima, Takashi Sawada, Norimasa Takakuwa, Yasunari

Tumor research : experimental and clinical ( 札幌医科大学 )  33   15 - 26  1998年

　概要を見る

To elucidate whether protein phosphorylation is associated with the loca-lization of the tight junction protein occludin, we determined the changes of occludin protein expression in primary cultures of rat hepatocytes after treat-ment with the protein phosphatase inhibitors okadaic acid and vanadate. After 2 h of treatment with 1myu M okadaic acid or 5 mM vanadate, occludin immunoreactivity showing continuous lines in non-treated cells changed to a few spots on the plasma membrane. In western blots, broad bands above the occludin protein (65 kD) became conspicuous after treatment with okadaic acid and vanadate. We treated the same samples with alkaline phosphatase to examine whether the broad bands depended on the changes in the phos-phorylation states of occludin protein. The broad bands disappeared and the occludin was observed as a narrow band corresponding to 65 kD. Neither a significant change in the mRNA of occludin nor a change in the immunoreac-tivity of the tight junction associated protein, ZO-1, was observed after treatment with okadaic acid or vanadate. These results suggested that the phosphorylation of occludin is closely associated with localization of the protein in cultured hepatocytes and that protein phosphatase inhibitors affect the loca-lization of occludin but not ZO-1 on the plasma membrane.

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Different changes in expression and function of connexin 26 and connexin 32 during DNA synthesis and redifferentiation in primary rat hepatocytes using a DMSO culture system

TL Kojima, M Yamamoto, C Mochizuki, T Mitaka, N Sawada, Y Mochizuki

HEPATOLOGY ( W B SAUNDERS CO )  26 ( 3 ) 585 - 597  1997年09月

　概要を見る

In the present study, we determined in detail the changes of liver gap junctions, connexin 26 (Cx26), and connexin 32 (Cx32), during DNA synthesis and redifferentiation of hepatocytes in vitro. We used primary rat hepatocytes that expressed the liver gap junction proteins, which were cultured in the medium containing epidermal growth factor (EGF) with 2% dimethylsulfoxide (DMSO) and 10(-7) mol/L glucagon (a DMSO culture system), as we previously reported, In the present cultures, almost confluent hepatocytes cultured in the medium containing EGF with 2% DMSO and 10(-7) mol/L glucagon, underwent a nearly synchronous wave of DNA synthesis induced by the removal of 2% DMSO and 10(-7) mol/L glucagon, and the addition of 10 mmol/L nicotinamide, after which the DNA synthesis was completely re-inhibited by the re-addition of 2% DMSO and 10(-7) mol/L glucagon, During stimulation of DNA synthesis, both Cx26 and Cx32 messenger RNA (mRNAs) in hepatocytes transiently increased in the G1 phase and then markedly decreased before the onset of the S phase, while only Cx26 messenger RNA (mRNA) increased slightly in the S/M phase. Furthermore, before the onset of the S phase, a disappearance of both Cx26 and Cx32 immunoreactivities and gap junction plaques were observed, Gap junctional intercellular communication (GJIC), as measured by lucifer yellow, which indicated the function of Cx32, decreased markedly from before the onset of the S phase, GJIC measured by propidium iodide, which indicated the function of Cx26, decreased from before the onset of the S phase and then increased slightly in the S/M phase. During the re-inhibition after the stimulation of DNA synthesis, Cx32 mRNA, but not Cx26 mRNA, rapidly returned to the pretreatment control level. Cx32 immunoreactivity and gap junction plaques also recovered. However, the recovery of GJIC measured by lucifer yellow was later than that of Cx32 expression. These results indicated the different changes of expression and function of Cx26 and Cx32 in the hepatocytes during stimulation and re-inhibition of DNA synthesis. This culture system should be useful as a model in which to study liver gap junctions during hepatocyte growth and differentiation in vitro.

DOI PubMed CiNii

Effects of melatonin on proliferation, oxidative stress and Cx32 gap junction protein expression in primary cultures of adult rat hepatocytes

T Kojima, C Mochizuki, T Mitaka, Y Mochizuki

CELL STRUCTURE AND FUNCTION ( JAPAN SOC CELL BIOLOGY )  22 ( 3 ) 347 - 356  1997年06月

　概要を見る

Melatonin has antiproliferative and antioxidant effects on cells in vivo and in vitro. Gap junctions mediate the communication between adjacent cells and are closely related to cellular growth and oxidative stress. We previously reported that 2% dimethylsulfoxide (DMSO) which has antiproliferative and antioxidative effects on hepatocytes, induces connexin 32 (Cx32) gap junction protein in primary cultures of adult rat hepatocytes. In the present study, we have examined the effects of melatonin on proliferation, oxidant stress and Cx32 gap junction protein expression in the cultured rat hepatocytes as compared to 2% DMSO treatment. 10(-2) M melatonin significantly inhibited the proliferation and the oxidative stress of the cells, and markedly induced Cx32 gap junction protein expression and gap junctional intercellular communication (GJIC). These effects of 10(-2) M melatonin treatment were not due to cytotoxicity to the cultured rat hepatocytes and they were as strong as those of 2% DMSO treatment. These results suggested that melatonin might be a useful substance to maintain the functions of the hepatocytes in vitro by modulating the levels of proliferation, oxidative stress and gap junction expression.

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Formation of actin filament networks in cultured rat hepatocytes treated with DMSO and glucagon

T Kojima, C Mochizuki, H Tobioka, M Saitoh, S Takahashi, T Mitaka, Y Mochizuki

CELL STRUCTURE AND FUNCTION ( JAPAN SOC CELL BIOLOGY )  22 ( 2 ) 269 - 278  1997年04月

　概要を見る

Actin filament organization may play an important role in the maintenance of differentiated functions in epithelial cells, We previously reported our success in inducing and maintaining gap junctions, which are two kinds of differentiated function, in primary rat hepatocytes cultured with 2% DMSO and 10(-7) M glucagon, In the present study, we demonstrated the formation of actin filament networks in the hepatocytes cultured with 2% DMSO and 10(-7) M glucagon, Actin filaments in hepatocytes cultured in medium with only 2% DMSO added from 96 h after plating were concentrated under the plasma membrane and were observed to be circumferential. In hepatocytes cultured in the medium with both 2% DMSO and 10(-7) M glucagon added from 96 h, not only the circumferential actin filaments but also the formation of actin filament networks were observed and the networks developed well with time in culture. The networks were observed as a dome-like structure under the cell face and terminated at the circumferential actin filaments, They were composed of electron-dense star-like vertices connected by microfilament bundles of varying length and were also very sensitive to the actin disrupter cytochalasin B, However, during the network formation, there were no significant increases in the amounts of actin protein and mRNA, The actin filament networks of the hepatocytes in this culture system might be closely related to the maintenance of differentiated functions.

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Expression of p21^wafl/cipl Is Induced in the Rat Liver by Feeding of 2-Acetylaminofluorene

Watanabe Tomoyuki, Ozaki Masakazu, Kawasaki Hajime, Kojima Takashi, Sawada Norimasa

Ｊｏｕｒｎａｌ ｏｆ Ｔｏｘｉｃｏｌｏｇｉｃ Ｐａｔｈｏｌｏｇｙ ( 日本毒性病理学会 )  11 ( 3 ) 191 - 196  1997年

　概要を見る

The cyclin-dependent kinase inhibitor p21^wafl/cipl plays major role in regulation of the cell cycle arrest following DNA damage in a p53-dependent and -independent manner. To elucidate the roles of p21^wafl/cipl in selective growth of preneoplastic lesions during chemical hepatocarcinogenesis, p21^wafl/cipl expression was examined in the livers of rats fed a 0.02% 2-acetylaminofluorene (2-AAF)-containing diet for 4 weeks or subjected to the protocol of Solt and Farber consisting of a single dose of diethylnitrosamine (DEN) followed 2 weeks later by 2-AAF feeding with two-thirds hepatectomy. By feeding of 2-AAF having a strong mitoinhibitory effect, p21^wafl/cipl expression was significantly induced in rat hepatocytes. The proliferating cell nuclear antigen (PCNA) labeling index of hepatocytes was consistently and significantly decreased by 2-AAF feeding. Immunohistochemically, both p21^wafl/cipl and p53 were detected in the livers of 2-AAF-fed rats more intensely than in those of control rats. In the livers of rats subjected to the protocol of Solt and Farber, most enzyme-altered foci (EAF) were immunohistochemically negative for p21^wafl/cipl, while hepatocytes surrounding the foci were positive. These results clearly show that 2-AAF acts on hepatocytes as a mitoinhibitor by inducing p21^wafl/cipl expression, presumably in a p53-dependent manner. Furthermore, the marked difference in p21^wafl/cipl expression between non-neoplastic and preneoplastic hepatocytes led to selective growth of preneoplastic lesions during 2-AFF feeding using the protocol of Solt and Farber.

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