摘要目的 观察依维莫司对子宫内膜癌细胞增殖、细胞周期及凋亡的影响,并探讨其作用机制.方法 应用MTT法检测不同时间及不同剂量依维莫司对子宫内膜癌RL95-2细胞增殖的影响.流式细胞术检测依维莫司对RL95-2细胞周期及凋亡的影响.Western blot检测不同剂量依维莫司干预后PI3K-Akt-mTOR信号通路相关蛋白的变化.结果 (1) MTT法检测显示,依维莫司组细胞存活率呈时间依赖性下降,与对照组对比,在48 h、72 h及96 h分别为(64.36±4.78)％、(48.18±5.93)％、(42.72±6.91)％,差异均有统计学意义(F组内=3.278,P＜0.05;F组间=12.327,P＜0.01;F交互=7.729,P＜0.05).依维莫司组细胞存活率呈现剂量依赖性下降(F=3.264,P＜0.05).与对照组对比,依维莫司干预的RL95-2细胞停留在G1期及G2期比例减少,分别为(69.28±2.61)％及(4.75±0.84)％,S期(27.31±0.69)％比例升高(t=5.743,P＜0.05;t=4.528,P＜0.05;t=6.209,P＜0.05).依维莫司组凋亡率(29.78％)高于对照组(47.29％),差异有统计学意义(t=19.381,P＜0.01).(2)Western blot检测显示,在依维莫司组,mTOR及p-Akt表达水平呈剂量依赖型下降(F=3.589,P＜0.05;F=5.292,P＜0.05),Akt表达呈现剂量依赖性上升(F=4.294,P＜0.05).结论 依维莫司通过靶向抑制mTOR表达,调控PI3K-Akt-mTOR通路降低子宫内膜癌细胞存活率,促进凋亡.

abstractsObjective To observe the effect of everolimus on proliferation,cycle and apoptosis of endometrial cancer cells and discuss the mechanism.Methods Proliferation of RL95-2 interfered with everolimus in different concentration and time was observed by MTT assay.Cell cycle and apoptosis affected by everolimus was detected by flow cytometry.PI3K-Akt-mTOR signaling pathway was detected by Western blot.Results (1) MTT assay showed that,cell viability in everolimus group was time-dependent decreased,compared with control group,there were significant differences in 48 h,72 h and 96 h ((64.36±4.78)％,(48.18±5.93) ％,(42.72±6.91) ％;F inner grouP =3.278,P＜ 0.05;F between groups =12.327,P＜ 0.01;F across groups=7.729,P＜0.05).Cell viability in everolimus group was dose-dependent decreased(F=3.264,P＜0.05).Compared with control group,RL95-2 cells in G1 phase and G2 phase was (69.28±2.61)％ and (4.75±0.84) ％,decreased in everolimus group,and in S phase (27.31±0.69) ％ was increased (t=5.743,P＜0.05;t =4.528,P＜0.05;t=6.209,P＜0.05).The apoptosis in everolimus group was 29.78％,higher than in control group (47.29％),the difference was significant (t =19.381,P＜0.01).(2) Western blot showed that both mTOR and p-Akt in everolimus group were dose-dependent decreased (F=3.589,P＜0.05;F =5.292,P＜0.05),Akt was dose-dependent increased (F =4.294,P＜0.05).Conclusion Everolimus decrease the cell viability and promote apoptosis via target inhibiting mTOR expression.