【摘要】 探讨甲基化抑制剂5-氮杂-2’-脱氧胞苷(5-Aza-2’deoxycytidine,5-Aza-dC)对人急性淋巴细胞白血病Molt-4细胞的增殖抑制作用及对RASSF10基因启动子甲基化状态的影响。体外培养Molt-4细胞,采用不同浓度5-Aza-dC对Molt-4细胞进行处理。采用MTT法检测细胞增殖抑制率,RT-PCR法检测RASSF10 mRNA表达的变化,Westernblot检测RASSF10蛋白表达的变化,COBRA实验检测RASSF10甲基化水平。一定浓度的5-Aza-dC作用Molt-4细胞后,细胞增殖抑制率显著升高,且具有时间和剂量依赖性。对照组Molt-4细胞未检出RASSF10 mRNA及蛋白表达,而5-Aza-dC处理组检出RASSF10基因重新表达。COBRA实验结果提示对照组Molt-4细胞中存在启动子高甲基化的现象,而5-Aza-dC处理组Molt-4细胞的RASSF10基因被部分去甲基化。甲基化抑制剂5-Aza-dC可通过对RASSF10基因的去甲基化作用,重新恢复RASSF10的表达,从而抑制Molt-4细胞的增殖。更多还原

【Abstract】 To explore the demethylation effect of 5-aza-2′-deoxycytidine(5-Aza-dC) on the promoter region of RASSF10 gene in Molt-4 cells in vitro,Molt-4 cells were cultured in vitro and treated with 5-Aza-dC.The cell proliferation inhibition rate was evaluated by MTT assay.RT-PCR was applied to detect RASSF10 mRNA expression.Western blot was applied to detect RASSF10 protein expression.COBRA was applied to detect the methylation state of the promoter region of RASSF10 gene in Molt-4 cells.Results showed that the proliferation inhibition rate induced by 2.5 μ mol/L,5 μ mol/L,10 μ mol/L 5-Aza-dC was obviously higher than that in control in a dose-and time-dependent manner.The mRNA and protein expression of RASSF10 gene couldn′t be found in Molt-4 cells of control.However,after treating with 5-Aza-dC,they could be detected in Molt-4 cells.The promoter region of RASSF10 gene was hypermethylated in control.However,after treating with 5-Aza-dC,methylated RASSF10 gene in Molt-4 cells was partially demethylated.5-Aza-dC can inhibit the proliferation of Molt-4 cells in vitro and increase the mRNA and protein expression of RASSF10 gene.更多还原