5-氮杂-2＇-脱氧胞苷对肺癌细胞H460生物学行为及抑癌基因RASSF1A表达的影响  

Effects of 5-Aza-2＇-deoxycytidine on biological behaviors and the RASSF1A gene expression of human lung carcinoma cell lines H460

作　　者：马红兵[1] 夏辉[2] 王西京[1] 

机构地区：[1]西安交通大学医学院第二附属医院肿瘤科,710004 [2]首都医科大学附属北京胸科医院国家结核病参比实验室

出　　处：《肿瘤研究与临床》2010年第7期469-472,共4页Cancer Research and Clinic

基　　金：基金项目：陕西省科技攻关项目（2004K13-G20）

摘　　要：目的 观察5-氮杂-2＇-脱氧胞苷（5-Aza-CdR）对肺癌细胞H460的生物学行为及RASSF1 A mRNA表达的影响.方法 5-Aza-CdR处理H460细胞,通过MTT方法、平板克隆试验观察细胞生长活性的变化,PCR检测RASSF1A甲基化状态,Western blotting法检测RASSF1A的蛋白表达,流式细胞术进行细胞周期分析.结果 H460细胞经5-Aza-CdR处理后,与未处理组相比,生长速度出现不同程度减慢,克隆形成率明显降低,RASSF1A甲基化程度降低,延缓H460细胞周期G1/S进程,使细胞阻滞于G1期.结论 在肺癌细胞系中,RASSF1A基因甲基化可能导致其表达缺失,而5-Aza-CdR能够恢复RASSF1A基因的表达,为肺癌的去甲基化治疗提供理论依据.Objective To investigate the effects of demethylating agent 5-Aza-2＇-deoxycytidine （5-Aza-CdR） on the biological behaviors and the expression of RASSF1A of lung carcinoma cells H460. Methods Lung carcinoma cell line H460 were treated with 5-Aza-CdR. Cell proliferation was determined by MTT assay and colony forming test. The methylation status of RASSF1A gene was detected by PCR. The expression of RASSF1A protein was measured by Western blotting, and the cell cycle was analyzed by flow cytometry. Results With 5-Aza-CdR treatment, the proliferation speed of H460 lung carcinoma cells was slowed down and the colony formation rate of H460 cells was decreased significantly compared with control group （38.5 %, 27.5 % and 60.5 % in 5-Aza-CdR 5 and 10 μmol/L groups and control group, respectively, P 〈0.05）. The methylation degree in the promoter of RASSF1A gene was decreased and the expression of RASSF1A protein was detected after 5-Aza-CdR treatment. 5-Aza-CdR induced G1 phase arrest of the H460 cells. Conclusion The hypermethylation of CpG island in the promoter of RASSF1A gene results in the loss of RASSF1A protein expression in human Lung carcinoma cell line. The demethylating agent 5-Aza-CdR could restore RASSF1A gene expression.These findings provide theoretic evidence for clinical treatment of human lung carcinoma with demethylation agent 5-Aza-CdR.

关 键 词：肺肿瘤 DNA甲基化 脱氧胞苷 

分 类 号：R734.2[医药卫生—肿瘤]