摘要
目的:观察哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)特异性小干扰RNA(mTOR-siRNA)干扰mTOR表达后,食管鳞癌EC9706细胞对雷帕霉素(rapamycin)敏感性的变化。方法:mTOR-siRNA转染EC9706细胞,RT-PCR检测干扰效果。mTOR-siRNA转染前后的EC9706细胞用雷帕霉素处理,Western blotting检测EC9706细胞中mTOR及其下游p70S6K蛋白的表达;流式细胞术检测EC9706细胞的周期及凋亡,CCK-8试剂盒检测EC9706细胞的增殖。结果:mTOR-siRNA下调EC9706细胞中mTOR mRNA的表达(P<0.05或P<0.01);雷帕霉素抑制EC9706细胞中mTOR和p-p70S6K蛋白的表达(P<0.05),并促进p70S6K蛋白表达(P<0.01),且mTOR-siRNA转染后此作用更明显(P<0.05)。雷帕霉素可诱导EC9706细胞凋亡(P<0.01)、抑制EC9706细胞增殖(P<0.05或P<0.01)、使EC9706细胞阻滞于G1期(P<0.01),且mTOR-siRNA转染后这些作用更强(P<0.05)。结论:mTOR-siRNA能特异性下调食管鳞癌EC9706细胞中mTOR表达,提高EC9706细胞对雷帕霉素的敏感性。
Objective：To investigate the sensitivity of esophageal squamous cell carcinoma EC9706 cells to rapamycin after silencing mTOR expression by small interfering RNA targeting mTOR（mTOR-siRNA）.Methods：EC9706 cells were transfected with mTOR-siRNA and the interference effect was investigated by RT-PCR.EC9706 cells were treated with rapamycin before and after mTOR-siRNA transfection,and the expressions of mTOR and its downstream p70S6K were detected by Western blotting analysis.Cell cycle,apoptosis and proliferation of EC9706 cells were determined by flow cytometry and CCK-8 kit,respectively.Results：mTOR-siRNA down-regulated the expression of mTOR mRNA in EC9706 cells（P〈0.05 or P〈0.01）.Rapamycin inhibited mTOR and phosphorylated p70S6K（p-p70S6K）expressions and increased p70S6K expression in EC9706 cells（all P〈0.05）,and these effects of rapamycin were further enhanced by mTOR-siRNA transfection（P〈0.05）.Rapamycin also induced apoptosis,inhibited proliferation and arrested cell cycle in G1 phase of EC9706 cells（all P〈0.01）,and transfection with mTOR-siRNA significantly promoted these effects of rapamycin in EC9706 cells（P〈0.05）.Conclusion：mTOR-siRNA can specifically down-regulate mTOR expression in esophageal squamous cell carcinoma EC9706 cells,and increase the sensitivity of EC9706 cells to rapamycin.
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2010年第4期392-397,共6页
Chinese Journal of Cancer Biotherapy
基金
国家自然科学基金资助项目(No.30901778)
郑州大学引进人才基金资助项目~~
