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目的:探讨5-氮杂胞苷对Hela宫颈癌细胞脆性组氨酸三联基因(Fragile histidine triad gene,FHIT)启动子去甲基化作用及对其增殖的影响。方法:采用低浓度和高浓度5-氮杂胞苷作用Hela宫颈癌细胞,分别采用BSP和RT-PCR检测处理前后FHIT基因启动子甲基化状态和mRNA表达,应用MTT法检测细胞增殖改变。结果:Hela宫颈癌细胞FHIT基因启动子表现高度甲基化状态(CG位点甲基化率为80%～100%),mRNA表达完全受到抑制,细胞呈指数增殖状态;低浓度5-氮杂胞苷作用后的甲基化率显著降低(10%～40%),细胞增殖速度降低;高浓度组FHIT基因启动子甲基化状态被完全逆转,mRNA表达明显高于低浓度组,细胞增殖速度显著低于正常组和低浓度组。结论:5-氮杂胞苷可逆转Hela宫颈癌细胞FHIT基因甲基化状态,使基因恢复表达而抑制细胞增殖,去甲基化和抑制增殖作用随5-氮杂胞苷剂量增加而增加。
Objective: To investigate the effect of 5-azacytidine on the demethylation and its proliferation of Fragile histidine triad gene (FHIT) promoter in Hela cervical carcinoma cells. Methods: Hela cervical cancer cells were treated with low and high concentrations of 5-azacytidine. The promoter methylation status and mRNA expression of FHIT gene were detected by BSP and RT-PCR respectively. Cell proliferation was detected by MTT assay. Results: The promoter of FHIT gene in Hela cervical cancer cells was highly methylated (the methylation rate was 80% -100% at CG site), the expression of FHIT gene was completely inhibited and the cells were exponentially proliferating. The low concentration of 5-aza The methylation rate of FHIT gene was significantly reduced (10% -40%) and the rate of cell proliferation was decreased. The methylation status of FHIT gene promoter was completely reversed in high concentration group, and the mRNA expression was significantly higher than that in low concentration group The speed was significantly lower than the normal group and low concentration group. CONCLUSION: 5-Azacytidine reverses the methylation status of FHIT gene in Hela cervical cancer cells, restores and regulates gene expression, and inhibits cell proliferation, demethylation and inhibition of proliferation as the doses of 5-azacytidine increase.