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(MTr)比色法检测细胞增殖,免疫印迹实验(Westernblot)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 A2.780 cells. Apoptosis was determined by flow cytometry ( FCM), MTr, and western blot as- say 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 in- volved with down-regulation of protein survivin.