脂质体介导端粒酶基因转染人口腔黏膜角化细胞的实验研究  

Transfecing hTRT into normal human oral keratinocytes by liposome

作　　者：周海文[1] 杨雯君[2] 王克敏[3] 周曾同[1] 

机构地区：[1]上海交通大学医学院附属第九人民医院·口腔医学院口腔黏膜科,上海200011 [2]上海交通大学医学院附属第九人民医院·口腔医学院口腔颌面外科,上海市口腔医学重点实验室,上海200011 [3]上海交通大学医学院生化教研室,上海200025

出　　处：《上海口腔医学》2009年第2期169-172,共4页Shanghai Journal of Stomatology

基　　金：国家自然科学基金(39970791);上海市教育委员会青年科研基金(04BC45)~~

摘　　要：目的:将端粒酶基因转染入人正常口腔黏膜角化细胞,观察人端粒酶反转录酶(hTRT)的表达情况。方法:采用脂质体LipofectAMINE 2000与转铁蛋白共同将含有hTRT基因的pEGFP-hTRT质粒转染人正常口腔黏膜角化细胞,经荧光显微镜、激光共聚焦显微镜观察荧光,并进行流式细胞仪检测转染效率,采用RT-PCR检测转染细胞中外源性hTRT的表达,并经G418筛选观察其稳定表达情况。结果:LipofectAMINE 2000与转铁蛋白共同将pEGFP-hTRT转染入人正常口腔黏膜角化细胞,转染率为15.34%,转染细胞表达外源性hTRT。结论:脂质体LipofectAMINE 2000与转铁蛋白成功介导含有端粒酶基因的pEGFP-hTRT质粒转染人口腔黏膜角化细胞。PURPOSE： To introduce human telomerase reverse transcriptase （hTRT） into normal human oral keratinocytes （OKC） to expand their life-span. METHODS： The pEGFP-hTRT was transferred into OKCs by lipofecAMINE2000 and transferring （TF）. Expression of hTERT in transfectantS was examined by fluorescence microscopy, laser confocal microscopy. Transfection efficiency was examined by flow cytometer. Exogenous hTERT gene was detected by RT-PCR. The transfeced cells were selected by G418 for stable expression. RESULTS： OKCs were transferred with pEGFP-hTRT by lipofecAMINE2000 and TF. The transfection efficiency was 15.34%. Exogenous hTERT gene could be defected by RT- PCR. The transfeced ceils exhibited transient expression of exogenous hTERT gene. CONCLUSIONS： The recombinant vector pEGFP-hTRT can be transfected into OKC by liposome. TF can improve transfection efficiency. Transient expression of exogenous hTERT is possible in oral keratinocytes. Supported by National Natural Science Foundation of China （Grant No.39970791）and Youth Science Foundation of Shanghai Municipal Education Commission （Grant No. 04BC45）.

关 键 词：端粒酶 脂质体 口腔黏膜角化细胞 pEGFP-hTRT质粒 基因转染 

分 类 号：R781.5[医药卫生—口腔医学]