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人DNA MTase反义基因转染对肝癌细胞DNA MTase基因mRNA表达的影响
引用本文:张宏,肖文华,梁后杰,房殿春,杨仕明,罗元辉. 人DNA MTase反义基因转染对肝癌细胞DNA MTase基因mRNA表达的影响[J]. 第三军医大学学报, 2002, 24(11): 1264-1266
作者姓名:张宏  肖文华  梁后杰  房殿春  杨仕明  罗元辉
作者单位:1. 第三军医大学附属西南医院肿瘤血液科,重庆,400038
2. 解放军第304医院肿瘤科,北京,100037
3. 第三军医大学附属西南医院全军消化专科中心,重庆,400038
基金项目:国家自然科学基金资助项目 ( 39870 34 4)
摘    要:目的:观察人DNA MTase反义基因转染对肝癌细胞系SMMC-7721内源性DNA MTase基因mRNA的表达的影响。方法:将将建的包含正、反义DNA MTase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-7721;采用RT-PCR法观察DNA MTase基因mRNA表达变化。结果:PCR法扩增外源性NeoR基因证实证用脂质体法成功将重组载体转染入细胞:RT-PCR法检测DNA MTase基因mRNA表达,显示转染反义DNA MTase基因片段的SMMC-7721细胞中DNA MTase基因mRNA表达下调。结论:人DNA MTase基因反义基因转染是一种有效,可靠的抑制肝癌细胞系SMMC-7721DNA MTase基因表达的方法。它为更多了解DNA MTase在肝癌发生的作用提供了帮助。

关 键 词:DNA甲基转移酶  真核表达载体  反义DNA  肝癌细胞系  mRNA
文章编号:1000-5404(2002)11-1264-03
修稿时间:2002-05-21

Effect of transfection of human antisense DNA MTase gene on expression of DNA MTase gene mRNA in SMMC-7721 cells line
ZHANG Hong,XIAO Wen hua,LIANG Hou jie,FANG Dian chun,YANG Shi ming,LUO Yuan hui. Effect of transfection of human antisense DNA MTase gene on expression of DNA MTase gene mRNA in SMMC-7721 cells line[J]. Acta Academiae Medicinae Militaris Tertiae, 2002, 24(11): 1264-1266
Authors:ZHANG Hong  XIAO Wen hua  LIANG Hou jie  FANG Dian chun  YANG Shi ming  LUO Yuan hui
Abstract: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.
Keywords:DNA methyltransferase  eukaryotic expression vector  antisense DNA  carcinoma  hepatocelluar  cell line  mRNA
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