慢病毒载体介导hTERT对肝细胞生物学特性的影响  

Influence on Biological Character Istics of Hepatocytes Transfected with hTERT Gene by Lentivirus

作　　者：郭伟[1] 李文媛[2] 

机构地区：[1]长治医学院附属和济医院普外二科,046011 [2]长治医学院药学系药理教研室

出　　处：《长治医学院学报》2011年第4期244-247,共4页Journal of Changzhi Medical College

摘　　要：目的:研究慢病毒介导人端粒酶逆转录酶(h-TERT)基因对人张氏肝细胞(chang liver)生物学特性的影响。方法:将表达h-TERT基因的重组慢病毒感染人张氏肝细胞,通过杀稻瘟筛选获得阳性克隆。分别采用实时荧光RT-PCR、TRAP-PCR及ELISA方法检测转染前后chang liver细胞的h-TERT-mRNA水平和端粒酶活性的变化。通过MTT法、流式细胞术(FCM)观察其形态变化及生长增殖情况。结果:转染后的chang liver细胞存在h-TERT基因过表达和端粒酶活性上调,体外培养条件下维持肝细胞原有形态,存活期显著延长,增殖能力较强。结论:外源性h-TERT基因的异位表达不仅上调chang liver细胞的端粒酶活性,而且促进其增殖。Objective：To investigate the influence on the biological characteristics of human hepatic cell line chang liver transfected with h-TERT gene mediated by lentiviral vector. Methods： The chang liver hepatocytes were infected with recombinant lentiviruses expressing h-TERT gene and the stable cells were selected by blastcidin. The h-TERT mRNA level was determined by Real-time PCR and the expression of telomerase activity was detected by TRAP-PCR and ELISA in the transfected cells. The new hepatocytes were observed and evaluated by molecular biological and morphologic methods such as MTT, flow cytometry. Results：The transfected chang liver cells had over expression of h-TERT gene and upregulated telomerase activity. They maintained original shape and exhibited prolonged life span in vitro. Conclusion： Eetopic expression of h-TERT in chang liver cells not only upregulates telomerase activity but also extends their replicative life span.

关 键 词：慢病毒 端粒酶 h-TERT基因 肝细胞 

分 类 号：R346[医药卫生—基础医学]