hTERT-TK／GCV对肝癌细胞生长及凋亡的靶向性影响  

Influence of hTERT-TK/GCV on growth and apoptosis of HCC

作　　者：杨长青[1] 邓志华[1] 王桂琴[2] 王晶晶[2] 曹燕[1] 

机构地区：[1]山西医科大学附属第二医院消化科,山西太原030001 [2]山西医科大学微免教研室,山西太原030001

出　　处：《中国癌症杂志》2008年第2期91-95,共5页China Oncology

基　　金：国家自然科学基金资助(No.30672405)

摘　　要：背景与目的:自杀基因治疗肿瘤已初见成效,引起了许多研究者的关注,端粒酶有希望成为肝癌基因治疗的理想靶点。本研究探讨hTERT-TK/GCV在体外对肝癌细胞生长及凋亡的靶向杀伤作用及机制。方法:体外细胞培养,构建荧光报告质粒和治疗质粒,脂质体瞬时转染方法将质粒导入肝癌细胞,荧光显微镜,流式细胞仪,原位末端标记等方法观察转染质粒对肝癌细胞生长及凋亡的影响,用Westernblot检测hTERT-TK对肝癌细胞周期调控因子cyclinD1、Cdk2、p21蛋白表达的影响。结果:hTERT-TK可以高效、特异的转染表达端粒酶活性的肝癌细胞HepG2,并使其凋亡,总的凋亡率可达64%,而正常肝细胞L-02的凋亡率仅为15%,几乎不影响正常肝细胞的生长。结论:hTERT-TK/GCV自杀基因系统具有靶向抗肝癌作用,有潜在临床应用前景。Background and purpose： The suicide gene therapy has attained preliminary effect in cancer gene therapy, more scientists have paid close attention to the strategy. Since telomerase was overactivated in many cancers, so it may serve as the ideal target in bepatoceilular carcinoma （HCC） gene therapy. We investigated the antitumor effect of the hTERT-TK/GCV system and its molecular mechanism in human HCC cell line HepG2 in vitro. Methods： Cells were cultured in vitro, luciferase reporter plasmid and therapeutic plasmid were constructed and transiently transfected into HepG2 ceils. The impacts of plasmid on ceil growth and apoptosis were estimated by luciferase microscope, flowcytometery and TUNEL assay. The cell-cycle modulator like cyclinD1, Cdk2 and p21 were determined by Western blot method. Results： HCC ceil line HepG2 with expression of telomerase activity could be transfected with pGL3-hTERT-TK plasmid more effectively and specifically. Transfected induced apoptosis was more profound in HepG2 than normal hepatic cell L-02 （64% vs 15%, P 〈0.05）. Conclusions： The hTERT-TK gene has the ability for targeted therapy in HCC, which provided a new strategy for clinical therapy against HCC.

关 键 词：靶向基因治疗 肝癌 HTERT启动子 自杀基因 

分 类 号：R731.31[医药卫生—肿瘤]