雷帕霉素对胃癌gc7901细胞增殖影响及其机制探讨.pdf

雷帕霉素对胃癌 SGC7901 细胞增殖影响及其机制探讨 中 文 摘 要 目的 研究雷帕霉素靶向抑制mTOR信号通路对胃癌SGC7901细胞增殖的影响 及其可能机制，为胃癌的药物治疗提供新的思路和方法. 方法 体外培养胃癌SGC7901细胞，用免疫细胞化学法检测缺氧条件下胃癌 SGC7901细胞中mTOR及HIF-1α蛋白表达情况。细胞经不同浓度雷帕霉素分别作用 12h、24h、48h、72h，CCK-8法检测雷帕霉素对细胞增殖的抑制作用。半定量RT-PCR 检测细胞mT0R和HIF-1αmRNA表达改变情况,Western Blot法检测细胞mT0R和 HIF-1α蛋白表达改变情况。实验所得数据采用SPSS13.0统计分析软件进行处理。 结果 免疫细胞化学显示，缺氧条件下胃癌SGC7901细胞胞浆中mTOR呈阳性 表达，细胞核和细胞浆中HIF-1α均呈阳性表达。雷帕霉素能显著抑制胃癌 SGC7901细胞增殖，抑制率随药物浓度和作用时间增加而增加，呈时间-剂量依赖 关系。雷帕霉素作用后，细胞mT0R和HIF-1αmRNA及蛋白表达水平均明显下调。 结论 缺氧条件下，胃癌SGC7901细胞存在mTOR/HIF-1α信号通路的异常活 化。在体外雷帕霉素能显著抑制胃癌SGC7901细胞增殖。缺氧条件下，雷帕霉素 抑制mTOR/HIF-1α信号通路是其可能机制之一。 关键词 雷帕霉素 胃肿瘤 哺乳动物雷帕霉素靶蛋白 缺氧诱导因子-1α 药物治疗 2 Effect of Rapamycin on human gastric carcinoma SGC7901 cells proliferation and approach of the mechanisms ABSTRACT Objective To investigate the effects of Rapamycin on cell proliferation in human gastric carcinoma SGC7901 cells and the potential mechanisms involved in the inhibitory effects of the mTOR signaling pathway. Provide the new ideas and original strategy about the drug treatment for human gastric carcinoma. Methods SGC7901 cells were cultured in vitro．The expression of mTOR and HIF-1αproteins was determined by immunocytochemical method in SGC7901 cells cultured under hypoxic conditions．Different concentrations of Rapamycin were incubated with SGC7901 cells for 12 hours、24 hours、 48 hours and 72 hours． Cell proliferation was measured by CCK-8 assay in Rapamycin—treated SGC7901 ce