Wang Huan, Li Dandan, Li Xiaomao, Ou Xueling, Liu Suiling, Zhang Yu, Ding Jie, Xie Bo
Department of Gynecology, Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong 510630, P.R. China.
Sun Yat-sen University Cancer Center, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Guangzhou, Guangdong 510060, P.R. China.
Oncol Lett. 2016 Dec;12(6):5029-5035. doi: 10.3892/ol.2016.5338. Epub 2016 Nov 2.
The aim of the present study was to investigate the effects of the mammalian target of rapamycin (mTOR) inhibitor, RAD001, on the growth of human endometrial cancer cells. The effects of RAD001 on human endometrial cancer Ishikawa and HEC-1A cell proliferation were determined by MTT assay. Green fluorescent protein microtubule-associated protein 1 light chain 3α (GFP-LC3) protein aggregates were observed under a confocal microscope, and Ishikawa and HEC-1A cell apoptosis was detected using flow cytometry. The expression levels of LC3-I, LC3-II and mTOR proteins were detected by western blot analysis. The results showed that RAD001 effectively inhibited human endometrial cancer Ishikawa and HEC-1A cell proliferation via downregulation of AKT/mTOR phosphorylation. Moreover, RAD001 induced autophagic cell death and a higher sensitivity to paclitaxel-induced apoptosis. These results indicate that RAD001 could have therapeutic potential in human endometrial cancer with hyperactivated AKT/mTOR signaling.
本研究的目的是探讨雷帕霉素靶蛋白（mTOR）抑制剂RAD001对人子宫内膜癌细胞生长的影响。采用MTT法测定RAD001对人子宫内膜癌Ishikawa细胞和HEC-1A细胞增殖的影响。在共聚焦显微镜下观察绿色荧光蛋白微管相关蛋白1轻链3α（GFP-LC3）蛋白聚集体，并用流式细胞术检测Ishikawa细胞和HEC-1A细胞凋亡。通过蛋白质印迹分析检测LC3-I、LC3-II和mTOR蛋白的表达水平。结果表明，RAD001通过下调AKT/mTOR磷酸化有效抑制人子宫内膜癌Ishikawa细胞和HEC-1A细胞增殖。此外，RAD001诱导自噬性细胞死亡，并对紫杉醇诱导的凋亡具有更高的敏感性。这些结果表明，RAD001在AKT/mTOR信号过度激活的人子宫内膜癌中可能具有治疗潜力。
