5Aza-dc对癌细胞TIMP-3启动子去甲基化的作用  

Effect of 5Aza-dc on demethylation of TIMP-3 gene promoter in carcinoma cells

作　　者：吕国丽[1,2] 文剑明[1] 张萌[1] 胥健敏[1] 徐若冰[1] 董愉[1] 

机构地区：[1]中山大学中山医学院病理学教研室,广东广州510080 [2]广州市刑事科学技术研究所

出　　处：《中国病理生理杂志》2005年第9期1763-1768,共6页Chinese Journal of Pathophysiology

基　　金：广东省自然科学基金资助项目(No.001315)

摘　　要：目的:观察5-氮杂-2’-脱氧胞苷(5-Aza-2’-deoxycytidine,5Aza-dc)对癌细胞TIMP-3启动子甲基化的影响。方法:用5Aza-dc处理TIMP-3启动子甲基化的H2M肝癌细胞和A431表皮癌细胞,用Transwell检测癌细胞的侵袭及运动能力,用Westernblot检测TIMP-3的蛋白表达,用RT-PCR检测TIMP-3mRNA的表达,用甲基化特异性PCR检测TIMP-3基因启动子的甲基化。结果:(1)5Aza-dc作用后的H2M和A431细胞的侵袭及运动能力降低;(2)5Aza-dc作用后的H2M和A431细胞的TIMP-3蛋白及mRNA表达量增加;(3)5Aza-dc作用后的H2M和A431细胞的TIMP-3启动子区未检测到甲基化。结论:5Aza-dc可以使肝癌和表皮癌细胞TIMP-3启动子区去甲基化,使TIMP-3得以重新表达,恢复其抑制肿瘤侵袭和移动的能力。AIM： To observe the effect of 5Aza - dc on demethylation of TIMP - 3 gene promoter in carcinoma cells. METHODS： Both hepatocellular carcinoma cells （H2M） and epidermoid carcinoma cells （A431） with methylation of TIMP- 3 promoter gene were treated with 5 - Aza - 2＇ - deoxycytidine （5Aza - dc）. Invasion ability and motility of the cells were detected by Transwell experiments. Expressions of TIMP - 3 protein and mRNA were detected by Western blotting and RT - PCR, respec- tively. TIMP-3 gene promoter methylation was detected by methylation- specific PCR （MSP）. RESULTS： （1） Invasion ability and motility of H2M and A431 cells were declined after treatment with 5Aza- dc; （2） After treatment with 5Aza - dc, the expression of TIMP- 3 protein and mRNA were increased in H2M and A431 cells; （3） After treatment with 5Aza- de, methylation of TIMP- 3 promoter gene was not detectable in the cell lines. CONCLUSIONS： 5Aza- de induces demethylation in TIMP- 3 promoter gene, restores TIMP- 3 gene and protein expression, and inhibits invasion ability and motihty of the carcinoma cells.

关 键 词：肝肿瘤 TIMP-3启动子 甲基化 脱氧胞苷 

分 类 号：R363[医药卫生—病理学]