白藜芦醇通过下调survivin蛋白表达促进人卵巢癌A2780细胞失巢凋亡的研究

摘要

目的:初步探讨人卵巢癌A2780细胞抗失巢凋亡的特性及白藜芦醇(RES)对人卵巢癌A2780细胞失巢凋亡的作用及相关机制.方法:利用悬浮培养模型体外模拟人卵巢癌A2780细胞失巢培养,流式细胞术(FCM)检测细胞凋亡；利用四甲基偶氮唑蓝(MTT)比色法检测细胞增殖,免疫印迹实验(Western blot)检测生存素(survivin)蛋白表达的情况.结果:悬浮培养A2780细胞凋亡率与正常贴壁培养细胞凋亡率相比[(3.19±1.63)％vs(3.51±1.33)％],差异无统计学意义(P＞0.05)；而悬浮培养组细胞survivin蛋白表达明显增高.RES处理后呈时间、剂量依赖性抑制悬浮培养A2780细胞增殖.RES处理组细胞凋亡率(58.71±6.78)％明显高于悬浮培养组细胞凋亡率(3.19±1.63)％(P＜0.01),且RES处理组细胞survivin蛋白水平明显下降.结论:人卵巢癌A2780细胞具有抗失巢凋亡的特性.RES能诱导人卵巢癌A2780细胞失巢凋亡,其效应与survivin蛋白表达下调有关.%Objective:To explore the effects of resveratrol (RES) on anoikis resistance and involved molecular mechanisms in human ovarian cancer A2780 cells.Methods:Suspension culture model was used to induce anoikis of A2780 cells.Apoptosis was determined by flow cytometry(FCM),MTr,and western blot assay were used to determine the cell viability and survivin protein expression respectively.Results:There was no significant difference in apoptotic ratio between cells cultured in suspension culture model and those in normal attached model [(3.19 ±1.63)％vs(3.51 ±1.33)％,P＞0.05].The survivin protein expression in cells under suspension culture model was obviously elevated.RES inhibited cell proliferation of A2780 cells under suspension culture model dose-and time-dependently.The apoptotic ratio of cells with RES treatment (58.71 ±6.78)％ was significantly higher than that of cells under suspension culture model (3.19 ± 1.63)％ (P＜0.01).The survivin protein level was obviously decreased in the cells treated with RES.Conclusions:The ovarian cancer A2780 cell is anoikis resistant.RES can induce anoikis of A2780 cells which may be involved with down-regulation of protein survivin.