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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、发展中的作用提供了帮助
Objective To observe the effect of DNAMTase antisense gene transfection on the expression of endogenous DNAMTase gene mRNA in hepatocellular carcinoma cell line SMMC 772 1. Methods The constructed eukaryotic expression vector containing sense and antisense DNAMTase gene fragments was transfected into hepatoma cell line SMMC 772 1 by lipofectamine. The mRNA expression of DNAMTase gene was observed by RT PCR. Results PCR amplification of exogenous NeoR gene confirmed that the recombinant vector was successfully transfected into cells by liposome. The mRNA of DNAMTase gene was detected by RT PCR. The results showed that DNAMTase gene was transfected into SMMC 772 1 cells transfected with antisense DNAMTase gene Downregulation of mRNA expression. Conclusion Human DNAMTase antisense gene transfection is an effective and reliable method to inhibit the gene expression of hepatocellular carcinoma cell line SMMC 772 1 DNAMTase. It helps to understand more about the role of DNAMTase in the occurrence and development of liver cancer