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。
AIM： To explore the effect of mammalian target of rapamycin （mTOR） inhibitor RAD001 on chemotherapeutic sensitivity of endometrial carcinoma ceils to paclitaxel. METHODS ： MTT assay and PI staining were used to assess the cell death. The protein expression of LC3-Ⅰ, LC3-Ⅱ, roTOR 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 ceils. RAD001 induced autophagy and autophagic cell death by the inhibition of roTOR/ 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.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2013年第11期1966-1971,共6页
Chinese Journal of Pathophysiology
基金
广州市科技计划资助项目(No.2012KP072)
