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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 the effect and mechanism of 5,7-dimethoxyflavone (5,7-DMF) on apoptosis in human breast cancer cell line MDA-MB-453. Methods: Breast cancer MDA-MB-453 cells were cultured in vitro. Cell viability was measured by MTT assay. Apoptosis rate was analyzed by flow cytometry (FCM) with propidium iodide (PI) staining. Methylation-specific polymerase chain reaction (MSP) was used to observe the methylation status of 14-3-3σ ; 14-3-3σ mRNA expression was detected by RT-PCR and 14-3-3σ protein was analyzed by Western blotting. Results: 5,7-DMF inhibited MDA-MB-453 cell activity and induced apoptosis in a concentration-dependent manner. Treatment with 10μmol / L 5,7-DMF for 72 hours significantly inhibited 14-3-3σ gene methylation and up-regulated its mRNA expression in MDA-MB-453 cells. 5,7-DMF up-regulates 14-3-3σ protein expression in a concentration-dependent manner. CONCLUSION: 5,7-DMF can induce the apoptosis of MDA-MB-453 cells by inhibiting the methylation of 14-3-3σ gene and up-regulating the expression of 14-3-3σ protein.