Misc - TOKINO Takashi

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

佐々木泰史, 今井浩三, 時野隆至

日本癌治療学会誌   37 ( 2 )  2002

J-GLOBAL

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

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

日本癌学会総会記事   61st  2002

J-GLOBAL

【ゲノム医療と情報システム】 情報システムのポストゲノムプロジェクトへの応用

辰巳 治之, 青木 文夫, 明石 浩史, 宮司 正道, 豊田 実, 佐々木 泰史, 西森 博幸, 時野 隆至, 伊東 文生, 今井 浩三

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

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

青木 文夫, 明石 浩史, 豊田 実, 宮司 正道, 佐々木 泰史, 西森 博幸, 時野 隆至, 辰巳 治之

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

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

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

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

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

DOI

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

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

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

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

DOI

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

佐々木 泰史, 時野 隆至

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

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

青木 文夫, 辰巳 治之, 明石 浩史, 宮司 正道, 豊田 実, 佐々木 泰史, 西森 博幸, 時野 隆至

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

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

D Kobayashi, T Tokino, N Watanabe

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

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

Mutational analysis of the beta-catenin gene in gastric carcinomas

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

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

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

DOI

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

佐々木泰史, 時野隆至

放射線生物研究   36 ( 3 )  2001

J-GLOBAL

潰よう性大腸炎におけるメチル化の異常と発癌リスク予測への応用

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

日本癌学会総会記事   60th  2001

J-GLOBAL

p53標的遺伝子としてのオステオポンチン遺伝子の同定

森本一郎, 佐々木泰史, 石田勢津子, 伊東文生, 今井浩三, 時野隆至

日本癌学会総会記事   60th  2001

J-GLOBAL

p53ノックアウトマウスを用いたp53標的遺伝子の同定

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

日本癌学会総会記事   60th  2001

J-GLOBAL

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

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

日本癌学会総会記事   60th  2001

J-GLOBAL

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

佐々木泰史, 坂本裕史, 時野隆至

日本消化器病学会雑誌   98  2001

J-GLOBAL

p53類似遺伝子p73,p63/p51の標的遺伝子の検索

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

日本癌学会総会記事   60th  2001

J-GLOBAL

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

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

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

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

DOI PubMed CiNii

子宮内膜癌におけるSRCの変異

小林 寛治, 寒河江 悟, 杉村 政樹, 西岡 善宏, 時野 隆至, 工藤 隆一

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

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

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

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

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