摘要
目的:通过观察应用去甲基化药物zebularine(Zeb)后人卵巢癌细胞系A2780中抑癌基因RASSF1A甲基化状态及表达水平的改变,探讨Zeb对卵巢癌细胞RASSF1A基因表达的调控。方法:应用不同浓度(50、100、200μmo.lL-1)的Zeb处理体外培养的卵巢癌细胞A2780后,甲基化特异性PCR(MSP)法检测用药前后细胞中RASSF1A基因的甲基化状态,RT-PCR法及Western-blotting法检测用药前后细胞中RASSF1A基因mRNA及蛋白表达的变化。结果:RASSF1A基因在卵巢癌细胞A2780中启动子区呈异常甲基化状态,在mRNA及蛋白水平表达阴性。经过Zeb处理后,RASSF1A基因启动子区呈去甲基化状态,其mRNA及蛋白重新表达。结论:去甲基化药物Zeb能使卵巢癌细胞RASSF1A基因呈去甲基化状态,从而调控RASSF1A基因重新表达。
Objective： Investigate the effect of zebularine（Zeb） on methylation and expression of RASSF1A gene in human ovarian cancer cell line A2780 and to discuss the regulating effect of demethylation agent on its expression.Methods： A2780 cells were cultured in RPMI 1640 and were treated with different concentrations（50,100,200 μmol·L-1） of DNA methyltransferase inhibitor Zeb.Methylation-specific PCP（MSP） was used to detect promoter methylation of RASSF1A gene.RT-PCR and Western blotting were used to detect expression of mRNA and protein of RASSF1A gene before and after treatment with Zeb respectively.Results： Promoter hypermethylation of RASSF1A gene was detected in ovarian cancer cell A2780,and no expression was detected before treatment.After treated with Zeb,the promoter region of RASSF1A gene exhibit demethylation,and its expression occured at mRNA and protein level.Conclusion： Zeb can induce demethylation of RASSF1A gene in human ovarian cancer cell A2780 and regulate its reexpression.
出处
《东南大学学报（医学版）》
CAS
2011年第4期550-554,共5页
Journal of Southeast University(Medical Science Edition)
基金
辽宁省教育厅科学基金资助项目(2008839)
辽宁省自然科学基金资助项目(20092116)
