5-Aza-dC对Molt-4细胞RASSF10基因启动子的去甲基化作用  

The demethylation effect of 5-Aza-2′ deoxycytidine on the promoter region of RASSF10 gene in Molt-4 cells in vitro

作　　者：汪茗[1] 章尧[1] 谢向荣 刘善浩 戚之琳[1] 毕富勇[1] 

机构地区：[1]皖南医学院生化教研室,芜湖241002 [2]弋矶山医院心内科,芜湖241001 [3]弋矶山医院血液内科,芜湖241002

出　　处：《生物学杂志》2013年第1期18-21,共4页Journal of Biology

基　　金：安徽省高校省级优秀青年人才基金项目(No.2011SQRL104);安徽省高校省级自然科学研究项目(No.KJ2011Z383);皖南医学院中青年科研基金资助项目(No.WK201010;No.WK2011F22)资助

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。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, Moh-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 expres- sion. COBRA was applied to detect the methylation state of the promoter region of RASSFIO gene in Moh-4 cells. Results showed that the proliferation inhibition rate induced by 2. 5 Ix mol/L, 5 Ix tool/L, 10 Ix 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 con- trol. However, after treating with 5-Aza-dC, they could be detected in Moh-4 cells. The promoter region of RASSF10 gene was byperm- ethylated in control. However, after treating with 5-Aza-dC, methylated RASSF10 gene in Moh-4 cells was partially demethylated. 5- Aza-dC can inhibit the proliferation of Moh-4 cells in vitro and increase the mRNA and protein expression of RASSF10 gene.

关 键 词：去甲基化 白血病 5-氮杂-2’-脱氧胞苷 RASSF10基因 

分 类 号：R733.7[医药卫生—肿瘤]