人DNA MTase反义基因转染对肝癌细胞DNA MTase基因mRNA表达的影响

Effect of transfection of human antisense DNA MTase gene on expression of DNA MTase gene mRNA in SMMC-7721 cells line

目的 观察人DNAMTase反义基因转染对肝癌细胞系SMMC 772 1内源性DNAMTase基因mRNA表达的影响。方法 将构建的包含正、反义DNAMTase基因片段的真核表达载体用脂质体法将其转染入肝癌细胞系SMMC 772 1 ;采用RT PCR法观察DNAMTase基因mRNA表达变化。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 ;RT PCR法检测DNAMTase基因mRNA表达 ,显示转染反义DNAMTase基因片段的SMMC 772 1细胞中DNAMTase基因mRNA表达下调。结论 人DNAMTase反义基因转染是一种有效、可靠的抑制肝癌细胞系SMMC 772 1DNAMTase基因表达的方法。它为更多了解DNAMTase在肝癌发生。

Objective To investigate the effect of the transfection of human antisense DNA MTase gene on the expression of ectogenetic DNA MTase gene mRNA of the human 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 by liposome DOTAP. The changes of the expression of DNA MTase gene mRNA were observed by RT PCR. Results The constructed recombinant vectors were successfully transfected into SMMC 7721 cell line with liposome DOTAP and affirmed by amplifying the ectogenetic NeoR gene with PCR. Decrease of the DNA MTase gene mRNA was found in the SMMC 7721 cell line tranfected with antisense DNA MTase gene fragment after being detected with RT PCR. Conclusion Transfection of human antisense DNA MTase gene was a valid way to inhibit the expression of DNA MTase gene of the SMMC 7721 cell line may be helpful for the derstanding the role of DNA MTase in the hepatocarcinoma.