癌变·畸变·突变2017，Vol.29Issue(4)：266-271,6.DOI:10.3969/j.issn.1004-616x.2017.04.005
一种转导人原代肝细胞的LV-HBx诱导调控表达系统
An inducible expression system of LV-HBx for transducing primary human hepatocyte
摘要
Abstract
OBJECTIVE:To construct an inducible lentiviral system which expressed HBx in primary human hepatocytes (PHH) for the study of biological characters of HBx and its influence on PHH.METHODS:We constructed a Tet-on inducible HBx-lentiviral expression system,and optimized it to a relatively high expression level. We applied this system to make LV-HBx and LV-rtTA recombinant lentivirus,to transduce HepG2 hepatoma cells and PHH,and to induce expression of HBx with doxycycline.RESULTS:We successfully constructed the recombinant lentiviral vector of HBx. The ratio of recombinant lentivirus of LV-HBx and LV-rtTA was optimized and a higher transduction efficiency could be approached at the ratio of 1:1 or 2:1 with HBx-positive cell ratio of 67.3% and 66.7%,respectively. This system was successfully applied in transducingHBx gene and inducing HBx expression with doxcycline in HepG2 and PHH. The expression level of HBx in PHH with Dox induction was about 45 fold of the group without Dox induction. HBx significantly increased the expression level of MTA1,VEGFA,and TGFB1 and decreased the expression level of CDH1 and IGFBP3,which is consistent with the previous reports.CONCLUSION:We successfully constructed a Tet-on expression system of LV-HBx for the transduction of HBx into PHH. This system will facilitate studies on influence of HBx and other HBV gene on PHH.关键词
HBx/人原代肝细胞/诱导表达/慢病毒载体Key words
HBx/primary human hepatocyte/inducible expression/lentiviral vector分类
生物科学引用本文复制引用
周小玲,熊小芳,谢庆东,孙平楠..一种转导人原代肝细胞的LV-HBx诱导调控表达系统[J].癌变·畸变·突变,2017,29(4):266-271,6.基金项目
国家自然科学基金(81570567,81571994) （81570567,81571994）
广东省自然科学基金(2014A030313483,2015A030313447) （2014A030313483,2015A030313447）
教育部留学回国人员科研启动基金(第49批) （第49批）
汕头大学医学院李嘉诚基金 （）
