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目的 :Molt-4细胞应用5-氮杂-2’-脱氧胞苷进行处理,并观察期对Molt-4细胞RASSF10基因启动子甲基化状态的影响。方法 :于体外进行Molt-4细胞培养,并采用不同浓度5-氮杂-2’-脱氧胞苷处理。然后采用MTT法检测性别增殖抑制率,同时采用RT-PCR法检测RASSF10 m RNA表达情况。采用COBRA检测RASSF10甲基化水平;Westernblot法检测RASSF10蛋白表达情况。结果 :经检测发现,采用一定浓度5-氮杂-2’-脱氧胞苷作用于Molt-4细胞后,细胞增殖抑制率明显升高;且表现为剂量依赖性和时间性。然对照组Molt-4细胞R中未检测出RASSF10 m RNA、蛋白表达情况。然经5-氮杂-2’-脱氧胞苷处理后,RASSF10基因出现部分甲基化。结论 :临床应用5-氮杂-2’-脱氧胞苷处理Molt-4细胞,其可经对RASSF10基因去甲基化来恢复RASSF10表达,最终达到抑制Molt-4细胞增殖效果。
Objective: Molt-4 cells were treated with 5-aza-2’-deoxycytidine and the effect of RASSF10 promoter methylation status was observed in Molt-4 cells. Methods: Molt-4 cells were cultured in vitro and treated with different concentrations of 5-aza-2’-deoxycytidine. Then MTT assay was used to detect the rate of inhibition of genistein proliferation. At the same time, the expression of RASSF10 mRNA was detected by RT-PCR. The methylation level of RASSF10 was detected by COBRA. The protein expression of RASSF10 was detected by Western blot. Results: The results showed that the inhibition rate of 5-AZA-2’-deoxycytidine was significantly increased after treated with Molt-4 cells in a dose-dependent and time-dependent manner. However, RASSF10 m RNA and protein expression were not detected in control group Molt-4 cells. However, 5-aza-2’-deoxycytidine treatment, RASSF10 gene partial methylation. CONCLUSION: Molt-4 cells are treated with 5-AZA-2’-deoxycytidine clinically, which can restore the expression of RASSF10 by demethylating RASSF10 gene and eventually inhibit the proliferation of Molt-4 cells.