电穿孔法转染人端粒酶逆转录酶至许旺细胞的研究  

Electransfection with telomerase catalytic subunit （hTERT） enhances proliferative capacity of Schwann cells

作　　者：张诚[1] 程飚[1] 出晓军[1] 王会仁[1] 陈峥嵘[1] 

机构地区：[1]复旦大学附属中山医院骨科,上海200032

出　　处：《中华显微外科杂志》2010年第3期213-216,I0006,共5页Chinese Journal of Microsurgery

基　　金：2006复旦青年骨干;上海市卫生局科研基金（2007041）;973课题基金赞助（2005CB522604）

摘　　要：目的利用电穿孔技术，将人端粒酶催化亚基（hTERT）转染至sD大鼠许旺细胞，观察及检测外源性bTERT在许旺细胞内表达及对许旺细胞寿命及增殖能力的影响。方法从食管癌组织中提取hTERT RNA，构建pcDNA3．1-hTERT质粒，利用电穿孔技术，转染许旺细胞，PCR检测hTERT RNA，TRAP—PCR银染法端粒酶活性检测试剂盒检测端粒酶活性，PI-annexin V染色，流式细胞仪测第15代转染及未转染ADSCs细胞凋亡，并且绘制15代转染及未转染细胞传代次数及传代时间曲线。结果成功构建pcDNA3．1-hTERT质粒，转染后，许旺细胞内hTERT RNA表达，并且TRAP端粒酶活性检测结果为阳性，许旺细胞标志物S100阳性，转染后细胞传代能力较未转染细胞显著增强。结论成功构建hTERT-许旺细胞，为构建许旺细胞永生化细胞系打下基础。Objective To transdnct with human telomerase reverse transcriptase （hTERT） to Schwann cells via electransfection technique to prolong the life span and enhance the proliferative capacity of Schwann cells. Methods The hTERT RNA was derived from esophagus cancer tissue and pcDNA3.1- hTERT vector was built. With electransfection technique, we transfected vector into ADSCs. The PCR, TRAP-PCR, and PI-annexin V were tested to prove the expression of hTERT in ADSCs. Results After transduction the hTERT RNA in Schwann cells, TRAP-PCR test, PI-annexin V dying and S100 dying was positive. Conclusion Telomerase catalytic subunit（hTERT） is transduct into Schwann cells, and it can to enhance the prolifertative capacity.

关 键 词：端粒酶 许旺细胞 电穿孔 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]