5,7-DMF诱导14-3-3σ基因甲基化促进乳腺癌MDA-MB-453细胞凋亡的实验研究  

Research on the Promotion of Apoptosis by 5,7-DMF in Breast Cancer Cells through Inducing the Methylation of 14-3-3σ Gene

作　　者：刘胜姿[1] 谭宇婷[1] 刘英姿[1] 刘飞[1] 周源[1] 

机构地区：[1]湖南师范大学医学院,湖南长沙410013

出　　处：《肿瘤药学》2013年第2期92-95,共4页Anti-Tumor Pharmacy

基　　金：湖南省卫生厅科研项目资助(No.B2010-058)

摘　　要：目的探讨5,7-二甲氧基黄酮(5,7-DMF)对人乳腺癌细胞凋亡的影响及其可能机制。方法体外培养乳腺癌MDA-MB-453、MCF-7细胞和永生化人乳腺上皮HBL-100细胞,经5,7-DMF处理后,通过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)检测14-3-3σ基因甲基化状态;Westernblotting分析14-3-3σ蛋白表达水平。结果 5,7-DMF对MDA-MB-453细胞活性的抑制作用最强;不同浓度(5、10和20μmol·L-1)5,7-DMF作用48小时后,可显著诱导MDA-MB-453细胞凋亡,其细胞内14-3-3σ基因甲基化水平显著升高,而14-3-3σ蛋白表达水平显著降低。结论 5,7-DMF可诱导乳腺癌MDA-MB-453细胞凋亡,其机制可能与促进14-3-3σ基因的甲基化和降低其蛋白表达有关。Objective To investigate the effects of 5, 7-DMF on the apoptosis of human breast cancer cells and its possible mechanisms. Methods Human breast cancer MDA-MB-453 cells, MCF-7 cells and human immortalized mammary epithelial HBL-100 cells were cultured in vitro. After treated with 5, 7-DMF, the cell viability was measured by MTT assay, and the rate of apoptosis was detected by using flow cytometry with PI staining. The methylation status of 14-3-3σ gene was determined by methlation specific PCR. （MSP）. The expression of 14-3-3σ protein was analyzed by Western blotting. Results 5,7-DMF showed the strongest activity against MDA-MB-453 cells in vitro. After treated for 48 h, 5,7-DMF （5, 10 and 20/μmol.L^-1） markedly induced the apoptosis of MDA-MB-453 cells, and the level of methylated 14-3-3σ gene was significantly increased, while the expression of 14-3-3σ protein was significantly decreased. Conclusion 5,7-DMF could induce the apoptosis of breast cancer MDA-MB-453 cells. Its mechanism may be related with the induction of the methylation of 14-3-3σ gene and the decrease of the expression of 14-3-3σ protein.

关 键 词：乳腺癌 5 7-二甲氧基黄酮 凋亡 14—3—3σ 

分 类 号：R737.9[医药卫生—肿瘤]