摘要
目的:研究5,7-二甲氧基黄酮(5,7-DMF)诱导人乳腺癌MDA-MB-453细胞凋亡作用及机制。方法:体外培养乳腺癌MDA-MB-453细胞。MTT法测定细胞活力;碘化丙啶(PI)染色流式细胞术(FCM)分析细胞凋亡率;甲基化特异性聚合酶链反应(MSP)观察14-3-3σ基因甲基化状态;逆转录聚合酶链反应(RT-PCR)检测14-3-3σ基因mRNA表达水平;Western blotting分析14-3-3σ蛋白表达。结果:5,7-DMF以浓度依赖方式抑制MDA-MB-453细胞活性和诱导细胞凋亡。10μmol/L 5,7-DMF处理72小时能显著抑制MDA-MB-453细胞14-3-3σ基因甲基化并上调其mRNA表达水平。5,7-DMF以浓度依赖方式上调14-3-3σ蛋白表达。结论:5,7-DMF诱导MDA-MB-453细胞凋亡与其抑制14-3-3σ基因甲基化并上调14-3-3σ蛋白表达有关。
Objective To investigate whether 5,7-dimethoxyflavone(5,7-DMF) induces apoptosis of human breast cancer MDA-MB-453 cells and its mechanism.Methods Human breast cancer MDA-MB-453 cells were cultured in vitro.The cell viability was measured by MTT assay.The apoptotic rate was examined using flow cytometry with PI staining.The methylation status of 14-3-3σ gene was observed by methlation specific PCR(MSP).The 14-3-3σ gene mRNA expression level was determined by reverse transcription PCR(RT-PCR).The expression of 14-3-3σ protein was analyzed by Western blotting.Results 5,7-DMF efficiently inhibited the viability of MDA-MB-453 cells and induced cell apoptosis in a dose-dependent manner.10 μmol/L 5,7-DMF significantly inhibited the methylation level of 14-3-3σ gene and upregulated its mRNA expression in MDA-MB-453 cells after treatment for 72 h.5,7-DMF upregulated the expression of 14-3-3 sigma protein of MDA-MB-453 cells in a concentraion-dependent manner.Conclusion 5,7-DMF efficiently induces apoptosis of breast cancer MDA-MB-453 cells,and its mechanisms are involved in inhibition of 14-3-3σ gene methylation and upregulating its protein expression.
出处
《湖南师范大学学报（医学版）》
2011年第4期9-12,共4页
Journal of Hunan Normal University(Medical Sciences)
