摘要
目的：研究5-氮杂-2＇脱氧胞苷（5～Aza—CdR）对肝癌细胞中DNA甲基转移酶1（DNMT1）及上皮钙黏附素（E-cadherin）表达的影响。方法：免疫组织化学检测上皮钙黏附素在肝癌（26例）及癌旁组织（20例）中的表达。用终浓度10μmol／L的5-Aza-CdR处理培养的肝癌细胞株GQY-7703作为实验组，未处理细胞为对照组；DNMT1基因和上皮钙黏附素基因（CDH1）的转录产物及表达蛋白分别使用半定量RT-PCR和免疫印迹法检测；CDH1的甲基化状态使用甲基化特异性PCR（MSP）检测。结果：相比较癌旁组织，肝癌组织中上皮钙黏附素表达显著下调。5-Aza-CdR降低了GQY-7703细胞中DNMT1在转录水平和翻译水平的表达；CDH1CpG岛的甲基化水平降低；上皮钙黏附素mRNA和蛋白表达量明显增高，对电泳条带的半定量分析显示实验组和对照组之间的差异具有统计学意义。结论：5-Aza-CdR通过抑制DNMT1的表达，改变了CDH1基因的异常甲基化状态，进而恢复上皮钙黏附素的表达。
Objective： To investigate the effects of cytosine methylation inhibitor 5-Aza-2＇-deoxycytidine （5-Aza-CdR） on the expression of DNA methytransferase 1 （DNMT1） and E-cadherin in hepatocellular carcinoma cells. Methods： Expression of E- cadherin in hepatocellular carcinoma and adjacent nontumorous liver tissue was detected by immunohistochemistry firstly. And then, the cultured hepatocellular carcinoma cells GQY-7703 was treated with 10μmol/L 5-Aza-CdR （final concentration）. The transcripts and protein of DNMT1 genes and E-cadherin genes （CDH1） were detected by semi-quantitative duplex RT-PCR （reverse transcription PCR） and Western blotting, respectively. The methylation level in the CpG island region of CDH1 was analyzed by methylation-specific PCR （MSP）. Results： The expression of E-caderin was down-regulated significantly in hepato- cellular carcinoma. The expression of DNMT1 mRNA and protein in hepatocellular carcinoma ceils treated with 5-Aza-CdR was significantly less than that of control. DNA hypermethylation in CpG island of CDH1 gene was reversed and leaded to up-regulation of CDH1 transcripts. The difference between the control and treatment in the expression of mRNA and E-cadherin protein analyzed by Gel-Pro analyzer had statistical significance. Conclusion： 5-Aza-CdR treatment can lead to DNA demethylation of CDH1 by inhibiting the expressions of DNMT1, and then reversed the expressions of E-cadherin mRNA.
出处
《解剖学杂志》
CAS
CSCD
北大核心
2010年第3期328-332,共5页
Chinese Journal of Anatomy
基金
安徽省教育厅自然科学研究项目（2006kj124c）
