摘要
目的探讨5-氮杂-2脱氧胞苷(5-Aza-2-deoxycytidine,5-Aza-Cde)对顺铂耐药株肺癌A549/DDP细胞DNA修复相关基因hMLH1、MGMT基因启动子区DNA甲基化状态及其表达的影响。方法 A549细胞(A549细胞组)、A549/DDP细胞(A549/DDP组)、A549/DDP细胞5、10、20μmol/L 5-Aza-Cde组,分别采用甲基化特异性PCR检测细胞hMLH1、MGMT基因甲基化状态,反转录-PCR法检测用药前、后细胞hMLH1mRNA、MGMT mRNA表达情况。结果A549细胞组hMLH1基因呈非甲基化状态且高表达,MGMT呈部分甲基化状态且高表达,A549-DDP细胞组hMLH1、MGMT基因均呈高甲基化状态,mRNA表达下调,10、20μmol/L 5-Aza-Cde组hMLH1和MGMT均呈非甲基化状态;A549/DDP细胞组、5μmol/L 5-Aza-Cde组hMLH1mRNA与MGMT mRNA表达量均低于A549细胞组(P<0.05),20μmol/L 5-Aza-Cde组与A549细胞组比较差异无统计学意义(P>0.05);10μmol/L 5-Aza-Cde组MGMT mRNA表达量低于A549细胞组(P<0.05),hMLH1mRNA表达量与A549细胞组比较差异无统计学意义(P>0.05)。结论 hMLH1、MGMT基因甲基化修饰程度与mRNA表达有一定相关性,hMLH1、MGMT基因甲基化可能是肺癌A549细胞对顺铂耐药的因素之一。
Objective To investigate the effects of 5-Aza-2＇-deoxycytidine （5-Aza-Cde） on DNA methylation and expression of mismatchrepair bMLH1 and MGMT gene in the human lung cancer cell line A549/DDP. Methods The methylations of hMLH1 and MGMT genes were determined by methylation-specific PCR, and the expressions of hMLH1 mRNA and MGMT mRNA were determined by reverse transcription-PCR before and after treatment in A549 group, A549/DDP group, and 5, 10 and 20 ptmol/L 5-Aza-Cde A549/DDP groups. Results In A549 group, hMLH1 showed no methylation and was highly expressed, and MGMT showed partial methylation and was highly expressed. In A549/DDP group, hMLH1 and MGMT showed hypermethylation, and hMLH1 mRNA and MGMT mRNA were downregulated. In 10 and 20μmol/L 5-Aza-Cde groups, hMLH1 and MGMT showed no methylation. The expressions of hMLH1 mRNA and MGMT mRNA were lower in A549/DDP group and 5 μmol/L 5-Aza-Cde group that those in A549/DDP group （P〈.05）, and there were no significant differences between 20μmol/L 5-Aza-Cde group and A549 group （P〈0.05）. The MGMT mRNA expression was lower in 10 μmol/L 5-Aza-Cde group than that in A549 group （P〈0.05）, and there was no significant difference in hMLH1 mRNA expression between these two groups （P 〈0.05）. Conclusions The methylations of hMLH1 and MGMT are correlated with their mRNA expression. And hMLH1 and MGMT methylation maybe one of the drug resistance factors for A549 to DDP.
出处
《中华实用诊断与治疗杂志》
2014年第2期122-124,共3页
Journal of Chinese Practical Diagnosis and Therapy
基金
湖北医药学院优秀中青年科技创新团队资助项目(ZD2012015)
