Analysis of molecular mechanism for modification of p53 tumor suppressor protein by anti-cancer drugs

抗癌药物修饰p53抑癌蛋白的分子机制分析

• 批准号: 18580289
• 负责人: ARAI Katsuhiko
• 金额: $ 2.57万
• 依托单位: Tokyo University of Agriculture and Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18580289/
• 关键词: tumor suppressor gene; p53; microtubule; tubulin; cell culture; anti-cancer drug; vinca alkaloid; vincristine; チュブリン

The aim of this research project is to obtain the information about the effects of anti-cancer drugs, especially vinca alkaloid, which play as a inhibitor of polymerization of microtubules, on the function of p53 superfamily. Head investigator, K. Arai found that β-tubulin isotypes transformed during establishment of drug-resistance for vinca alkaloid in mouse melanoma cell line, B16F10. Normally, expression of-tubulin expression was suppressed by p53 protein and decrease of DNA binding activity of Sp1 and p53 was induced by addition of vinca alkaloid to the culture medium. In 2006, to examine whether vinca alkaloid affect interaction between p53 superfamily and Sp1, in vitro translation system for p53 superfamily and Sp1 was established. Briefly, full length cDNAs corresponding to transactivating forms for p63, p73 and Sp1 in addition to wild-type and mutated p53 were amplified, then subcloned into pcDNA3.1 or pcDNA3.1/V5-His and used for in vitro transcription/translation. These enable to analyze interaction between p53 superfamily and Sp1 and their kinetics. In 2007, forced expression of p53 superfamily genes in p53-deficient mouse macrophage cell line was designed. Furthermore, mRNA distribution of p53 superfamily in the lung was examined by in situ hybridization and ribonuclease protection assay. As a result, mRNA corresponding to transactivating form of p73 was expressed in bronchial and type II alveolar epithelium. These findings indicated that p73 participate lung carcinogenesis.

该研究项目的目的是获得有关抗癌药物，尤其是作为微管聚合抑制剂的长春花生物碱对 p53 超家族功能的影响的信息，首席研究员 K. Arai 发现 β。在小鼠黑色素瘤细胞系 B16F10 中，在建立长春花生物碱耐药性期间，β-微管蛋白同种型发生了转化。正常情况下，β-微管蛋白的表达为2006年，为了检测长春花生物碱是否影响p53超家族和Sp1之间的相互作用，p53超家族的体外翻译系统和Sp1被p53蛋白抑制并降低了Sp1和p53的DNA结合活性。简而言之，除了野生型和突变型之外，还建立了对应于p63、p73和Sp1的反式激活形式的全长cDNA。 p53 被扩增，然后亚克隆到 pcDNA3.1 或 pcDNA3.1/V5-His 中，并用于体外转录/翻译，这使得能够分析 p53 超家族和 Sp1 之间的相互作用及其动力学。 2007 年，p53 超家族基因的强制表达。设计了 p53 缺陷型小鼠巨噬细胞系。此外，通过原位杂交检查了 p53 超家族的 mRNA 分布。结果，对应于p73反式激活形式的mRNA在支气管和II型肺泡上皮中表达。这些结果表明p73参与肺癌发生。

项目成果

Active Expression of Matrix Metalloproteinase-13 mRNA in the Granulation Tissue of Equine Superficial Digital Flexor Tendinitis.

马浅指屈肌腱炎肉芽组织中基质金属蛋白酶 13 mRNA 的活性表达。

• 发表时间: 2007
• 期刊: J. Vet. Med. Sci. 69
• 作者: Nomura;et. al.
• 通讯作者: et. al.

Piperonyl butoxide activates c-Jun and ATF-2 in the hepatocytes of the liver of mice

胡椒基丁醚激活小鼠肝脏肝细胞中的 c-Jun 和 ATF-2

• 发表时间: 2008
• 期刊: Arch. Toxicol. (In press)
• 作者: Muguruma;et. al.
• 通讯作者: et. al.

Search for matrix degrading activity in a case of equine superficial digital flexor tendonitis

寻找马浅表屈肌腱炎病例中的基质降解活性

• 发表时间: 2007
• 作者: H.;Satoh;M.;Nomura;Y.;Kasashima;S.;Hattori;K.;Arai
• 通讯作者: Arai

神経特異的クラスIIIβ-チューブリン遺伝子上のレチノイン酸依存性制御領域の解析

神经元特异性 III 类 β-微管蛋白基因上视黄酸依赖性调节区的分析

• 发表时间: 2007
• 作者: 石井;ほか
• 通讯作者: ほか

ウマ浅指屈腱炎症例の腱組織におけるマトリックス分解活性の検索

寻找马浅屈肌腱炎病例肌腱组织中的基质降解活性

• 发表时间: 2007
• 作者: 佐藤;ほか
• 通讯作者: ほか