白藜芦醇对人子宫内膜癌细胞系AN3CA增殖和凋亡效应及其机制探讨

目的：探讨白藜芦醇（resveratrol，Res）对人子宫内膜癌细胞AN3CA的增殖抑制和凋亡诱导效应及可能存在的机制。方法：应用噻唑蓝（MTT）法检测Res对AN3CA的增殖影响；流式细胞术检测Res对细胞周期分布和凋亡影响：荧光实时定量PCR检测Res对细胞Bcl-2、Bax和MMP-9mRNA表达水平的影响；WesternBlot方法检测Res对PCNA、Bcl-2、Bax及ERK1／2、P—ERK1／2蛋白表达水平的影响。结果：Res对子宫内膜癌细胞AN3CA具有显著的生长抑制作用（P〈0．01），呈时间-剂量依赖性；不同浓度Res处理细胞G0／G1期比例显著增加伴随S期细胞数的减少；细胞凋亡率明显增高，200Dmol／lRes处理48h凋亡率可达30．96％±2．041％（P〈0．01）。与对照组相比，Res能抑制PCNA的蛋白表达量，增加Bax和降低Bcl-2转录和蛋白水平的表达量。Res在短时间内（0．5—1h）激活ERK1／2的磷酸化表达但随着作用时间延长（4—48h）其表现为抑制效应。结论：Res具有抑制AN3CA细胞增殖，诱导细胞G0／G1期阻滞和凋亡的效应。Res诱导凋亡可能是通过上调Bax，下调Bcl-2发挥作用，其抗癌作用机制可能与ERK1／2通路失调相关。

Effects of Resveratrol on Proliferation and Apoptosis of Endometrial Carcinoma AN3CA Cells and Its Mechanism

Objective： To investigate the effects of resveratrol （Res） on proliferation and apoptosis of endometrial cancer cell lines AN3CA and explore the involved mechanism in vitro. Methods： AN3CA cells were treated with different concentrations of resveratrol for various periods. The cell growth inhibitory rate was examined by MTT assay. The cell cycle arrest and apoptosis induction of resveratrol in AN3CA cells were detected by flow-cytometry analysis.We utilized real-time quantitative polymerase chain reaction（PCR） to identify expression level of Bcl-2,Bax and MMP-9 mRNA.The protein expression level of PCNA,Bcl-2,Bax,ERK1/2 and p-ERK1/2 were detected by Western-blot.Results： Resveratrol has significantly anti-proliferation effect on AN3CA cells which is in time- and concentration-dependent manners. It increases the G0/G1 phase cell population, whereas a concomitant decrease of cell numbers in S phase（P 〈0.01）. In addition, resveratrol treatment could induce cellular apoptosis that the rate of apoptosis increased to 30.96%±2.041% （200μmol/1 Res）.Compared with the control, resveratrol treatment caused a decrease in the levels of PCNA and Bcl-2,whereas the level of Bax expression was upregulated. In a short time （0.5-1h）, resveratrol can activate ERK 1/2 phosphorylation but with the time prolonged, it showed inhibition of the expression ofp-ERK. Conclusion： It seems that resveratrol has the antitumor action including inhibition of cell proliferation, blocking cells at the G0/G1 phase and inducing apoptosis. It regulated expression level of apoptosis-related Bcl-2 family protein. The antitumor effect ofresveratrol is possiblly mediated by ERK1/2 pathway.