分泌型凋亡素真核表达载体的构建及对肝癌细胞HepG2凋亡的影响  

Construction of eukaryotic expression vector of NT4-APOPTIN-HA2-TAT in secretory form and its effects on the tumor tells

作　　者：王健生 张明鑫[1] 刘德纯[1] 段小艺[1] 王全颖 杨广笑 

机构地区：[1]西安交通大学医学院第一附属医院,710061 [2]陕西华广生物公司

出　　处：《中华肝胆外科杂志》2009年第6期464-466,共3页Chinese Journal of Hepatobiliary Surgery

基　　金：基金项目：该课题受高等学校博士学科点专项科研基金（20060698055）,教育部新世纪优秀人才支持计划（NCET-060838）资助

摘　　要：目的构建含有NT4和HA2-TAT的分泌型VP3真核表达载体，并观察其表达对人肝癌细胞HepG2和小鼠成纤维细胞NIH3T3的影响。方法构建了含鸡贫血病毒VP3基因和NT4、HA2-TAT基因的分泌型真核表达载体——NT4-Apoptin—HA2-TAT重组腺相关病毒载体。体外转染人肝癌细胞HepG2和小鼠成纤维细胞NIH3T3，用MTT法和流式细胞学，检测凋亡素融合基因诱导细胞凋亡的效果。结果携带融合基因的重组腺相关病毒感染人肝癌HepG2细胞48h后，显示了很强的诱导肿瘤细胞凋亡的能力。而对小鼠成纤维细胞NIH3T3细胞无明显凋亡作用。结论分泌型NT4-ApoptinHA2-TAT重组腺相关病毒载体转染细胞后能够被正常分泌、穿膜并特异性诱导肿瘤细胞凋亡，为肿瘤的基因治疗提供了新的思路。Objective To explore antitumor effect of eukaryotic expression vector of NT4 Apoptin-HA2-TAT in secretory form. Methods Recombinant vector fusing with Apoptin （VP3）, NT4, HA2-TAT were constructed and transfected into HepG2 and NIH3T3 cells. The results of cell death were detected by MTT and flow cytometry. Results The recombined adeno-associated virus fu- sing NT4 Apoptin-HA2-TAT could induce apoptosis 48h after HepG2 cells were transfected. However, it was not seen in normal NIH3T3 cells. Conclusion NT4-Apoptin-HA2 TAT can be secreted to specifically induce apoptosis of tumor cells, which provides a new way of anti-tumor gene therapy.

关 键 词：癌 肝细胞 分泌 凋亡素 真核表达载体 肿瘤细胞凋亡 

分 类 号：R686[医药卫生—骨科学]