人DNA MTase反义基因转染对诱导肝癌SMMC7-721细胞凋亡的影响

Apoptosis of hepatocarcinoma SMMC-7721 cells 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 1凋亡的影响。方法 采用脂质体法将构建的包含正、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;采用MTT法绘制生长曲线 ;流式细胞法和原位末端标记法检测细胞凋亡改变。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 ;生长曲线显示转染反义DNAMTase基因片段的SMMC 772 1细胞增殖速度减慢 ;流式细胞法测得其凋亡率由 3 1%增加到 13 5 %。末端标记法也显示其凋亡指数增加。结论 人DNAMTase反义基因转染可诱导肝癌细胞系SMMC 772 1凋亡。

Objective To investigate the change of the apoptosis of the human hepatocarcinoma cell line SMMC 7721 transfected by antisense human DNA MTase gene fragment. Methods The constructed sense and antisense DNA MTase gene eukaryotic expression vectors were transfected into the hepatocarcinoma cell line SMMC 7721 with liposome DOTAP, growth speed was observed by MTT assay and the change of the apoptosis of SMMC 7721 cell line by flow cytometry and TUNEL method. Results The sense and antisense eukaryotic expression vectors were successfully constructed and affirmed by restriction endonuclease analyzing and sequencing. The constructed recombinant plasmids were successfully transfected into SMMC 7721 cell line with liposome DOTAP and affirmed by amplifying the ectogenetic NeoR gene with PCR. The growth curve showed lower growth speed in the SMMC 7721 cell line transfected with antisense DNA MTase gene fragment. The apoptosis rate increased from 3.1% to 13.5% and apoptosis index also increased. Conclusion Transfection of human antisense DNA MTase gene can induce the apoptosis of the SMMC 7721 cell. Recovery of the function of the apoptosis genes may be one of the main causes.