【摘要】 目的:探讨哺乳动物雷帕霉素靶蛋白(mTOR)抑制剂RAD001通过诱导细胞自噬增强紫杉醇杀伤子宫内膜癌细胞作用的机制。方法:用MTT法检测RAD001对人子宫内膜癌Ishikawa和HEC-1A细胞的生长抑制作用,用激光共聚焦显微镜观察GFP-LC3蛋白的聚集;用流式细胞术检测细胞死亡;用Western blotting方法检测LC3-I、LC3-II、mTOR及ULK1蛋白的表达;用靶向ULK1的siRNA特异性地抑制Ishikawa细胞中ULK1的表达,再检测相关指标。结果:RAD001可抑制Ishikawa和HEC-1A细胞的增殖,提高它们对紫杉醇的敏感性。RAD001诱导Ishikawa和HEC-1A细胞发生自噬及自噬性细胞死亡。RAD001通过抑制mTOR/p70S6K通路、上调ULK1诱导自噬,从而产生紫杉醇增敏作用。结论:RAD001可以通过抑制mTOR信号通路,上调ULK1的表达,诱导自噬性细胞死亡的发生,从而提高子宫内膜癌细胞对紫杉醇的敏感性。更多还原

【Abstract】 AIM: To explore the effect of mammalian target of rapamycin( mTOR) inhibitor RAD001 on chemotherapeutic sensitivity of endometrial carcinoma cells to paclitaxel. METHODS: MTT assay and PI staining were used to assess the cell death. The protein expression of LC3-I,LC3-II,mTOR and ULK1 was detected by Western blotting.ULK1 siRNA was used to abolish the activation of ULK1. RESULTS: RAD001 significantly inhibited the growth of human endometrial carcinoma cell lines Ishikawa and HEC-1A. RAD001 enhanced the inhibitory effect of paclitaxel on the growth of Ishikawa cells and HEC-1A cells. RAD001 induced autophagy and autophagic cell death by the inhibition of mTOR /p70S6K pathway and up-regulation of ULK1 expression. CONCLUSION: RAD001 enhances the inhibitory effect of paclitaxel on endometrial carcinoma cell growth by inducing autophagy and autophagic cell death.更多还原