丹参酮ⅡA诱导人卵巢癌SKOV3细胞凋亡及机制的研究

The Studies of Apoptosis Effect and Its Mechanisms of Tanshinone ⅡA on Human Ovarian Cancer Cell SKOV3

目的:观察中药丹参有效提取成分之一丹参酮ⅡA(TanⅡA)对人卵巢癌细胞株SKOV3的增殖和凋亡的诱导作用,及对基质金属蛋白酶-2(MMP-2)和血管内皮生长因子(VEGF)表达的影响,探讨其可能的抗肿瘤作用机制。方法:体外培养卵巢癌SKOV3细胞,随机分组,实验组给予不同浓度TanⅡA处理细胞,倒置显微镜及四甲基偶氮唑盐(MTT)比色法观察实验组与对照组细胞增殖抑制情况;流式细胞术检测凋亡率及细胞周期变化;免疫荧光细胞化学法检测细胞MMP-2、VEGF蛋白表达改变;实时荧光定量聚合酶链反应(RT-PCR)检测MMP-2、VEGFmRNA表达水平。结果:TanⅡA对人卵巢癌SKOV3细胞的增殖具有抑制作用,呈时间-剂量依赖性;实验组较对照组G0/G1期细胞数量增多,而S期细胞数量减少;MMP-2、VEGF蛋白表达及基因表达明显下调,与对照组相比差异有统计学意义(P<0.05)。结论:TanⅡA对卵巢癌SKOV3细胞有明显的增殖抑制及诱导凋亡作用;TanⅡA可下调细胞MMP-2、VEGF的表达,这可能是其抗肿瘤作用机制之一。

Objective：To investigate the inhibition function of Tanshinone ⅡA which extracts from Tanshinone on human ovarian cancer SKOV3 cell growth,and detect the expression of MMP-2 and VEGF in control group and the experimental group,then explore it＇s mechanism. Methods：SKOV3 cell was cultured in vitro and divided into 2 groups randomly, the experimental groups were treated by Tan IIA with different dose. Inverted microscope was used to detect apoptosis. The proliferation inhibition was examined by MTT assay,and cell cycles and apoptosis were examined by flow cytometry（FCM） .RT-PCR and immunocytochemical method were used to detect the expression of MMP-2 and VEGF. Results：Tan IIA could introduce apoptosis in human ovarian cancer SKOV3 cell, and the effect paralleled with time and the concentration of drug, acted as a time-dose-dependent manner.FCM show that after treated by Tan IIA for 24、48、72 h, the apoptotic rate increased . RT-PCR and immunocytochemical method detected the protein and mRNA expression of MMP-2 and VEGF declined in experimental group, and the difference was statistically significant （P0.05）. Conclusions： Tan IIA can significantly inhibit the proliferation of human ovarian cancer SKOV3 cells and induce the cell apoptosis. Declining the expression of MMP-2 and VEGF might be the mechanism that Tan IIA induced apoptosis in human ovarian SKOV3 cells.