人DNA MTase反义基因转染诱导E-Cadherin基因启动子区域去甲基化

Demethylation in the promoter region of E-Cadherin gene induced by the transfected human antisense DNA MTase gene

目的 观察人DNAMTase反义基因转染对肝癌细胞系SMMC 772 1E Cadherin基因启动子区域甲基化状态的影响。方法 采用脂质体法将已构建的包含正、反义DNAMTase基因片段的真核表达载体转染入肝癌细胞系SMMC 772 1;用甲基化特异的PCR法检测E Cadherin基因启动子区域甲基化状态的改变。结果 PCR法扩增外源性NeoR基因证实用脂质体法成功将重组载体转染入细胞 ;甲基化特异的PCR法检测显示转染反义DNAMTase基因片段的SMMC 772 1细胞中的E Cadherin基因启动子区域呈现去甲基化状态。结论 人DNAMTase反义基因片段可诱导E Cadherin基因启动子区域去甲基化 ;实验结果有助于进一步了解DNAMTase在肝癌发展中的作用。

Objective To investigate the effects of the transfection of human antisense DNA MTase gene on the methylation status in the promoter region of E Cadherin gene in hepatocarcinoma cell line SMMC 7721. Methods The constructed sense and antisense DNA MTase gene eukaryotic expression vectors were transfected into the hepatocarcinoma cell line SMMC 7721 with liposome DOTAP and the methylation status of E Cadherin gene in the promoter region was detected with methylation specific PCR(MSP). Results The constructed recombinant vectors were successfully transfected into SMMC 7721 cell line with liposome DOTAP affirmed by amplifying the ectogenetic NeoR gene with PCR. Demethylation with MSP was found in the promoter region of E Cadherin gene of the SMMC 7721 cell line tranfected with antisense DNA MTase gene fragment. Conclusion Human antisense DNA MTase gene can induce demethylation in the promoter region of E Cadherin gene, and maybe helpful for understanding the role of DNA MTase in the development of hepatocarcinoma.