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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 observe the effect of mammalian target of rapamycin (mTOR) specific small interfering RNA (mTOR-siRNA) on the expression of mTOR in esophageal squamous carcinoma cell line EC9706, which is sensitive to rapamycin Variety. Methods: EC9706 cells were transfected with mTOR-siRNA and the interference effect was detected by RT-PCR. The EC9706 cells before and after mTOR-siRNA transfection were treated with rapamycin, and the expression of mTOR and its downstream p70S6K protein in EC9706 cells was detected by Western blotting. The cycle and apoptosis of EC9706 cells were detected by flow cytometry and CCK-8 kit EC9706 cell proliferation. Results: mTOR-siRNA down-regulated the expression of mTOR mRNA in EC9706 cells (P <0.05 or P <0.01); rapamycin inhibited the expression of mTOR and p-p70S6K in EC9706 cells (P <0.05) (P <0.01), and this effect was more obvious after mTOR-siRNA transfection (P <0.05). Rapamycin could induce the apoptosis of EC9706 cells (P <0.01), inhibit the proliferation of EC9706 cells (P <0.05 or P <0.01), and block the EC9706 cells in G1 phase (P <0.01) After these effects stronger (P <0.05). Conclusion: mTOR-siRNA can specifically down-regulate mTOR expression in esophageal squamous cell carcinoma EC9706 cells and improve the sensitivity of EC9706 cells to rapamycin.