APOBEC3G体外抗乙型肝炎病毒的研究

Inhibition of hepatitis B virus replication by APOBEC3G in vitro

目的探讨载脂蛋白B mRNA编辑酶催化多肽样蛋白3G(apolipoprotein B mRNA editing enzyme catalytic polypeptide like3G,APOBEC3G)(也称为CEM15)体外抗乙型肝炎病毒(HBV)的作用及其机制。方法脂质体转染pcDNA3.1 Human APOBEC3G-Myc-6Xhis、pcDAN3.1/His-C进入HepG2.2.15细胞,转染后,RT-PCR证实转染基因的表达,Western Blot证实蛋白的表达。通过ELISA方法检测细胞上清液中乙型肝炎表面抗原(HBsAg)及乙型肝炎e抗原(HBeAg),RT-PCR分析APOBEC3G对HBV mRNA转录的影响。结果APOBEC3G基因与蛋白在HepG2.2.15细胞都有表达,与空质粒转染组相比,pcDNA3.1 Human APOBEC3G-Myc-6Xhis转染组HBsAg含量下降70.38%,HBeAg含量下降62.88%,未转质粒细胞为空白对照组。结论APOBEC3G在体外可以抑制HBV复制,可以作为一种新型的抗病毒制剂治疗乙肝病毒感染。

Objective To validate specific anti-viral effects and mechanism of apolipoprotenin B mRNA editing enzyme catalytic polypeptide like 3G（APOBEC3G） on replication and expression of hepatitis B virus in vitro. Methods After transfection of pcDNA3. 1 Human AP OBEC3G-Myc-6Xhis into HepG2.2. 15 cell line,the expression of APO- BEC3G were detected at mRNA and protein levels by RT-PCR and Western Bolt,respectively. ELISA and RT-PCR were applied to measure the alteration of expressions of HBsAg and HBeAg in supernatant of HepG2.2.15 cell lines. Simultaneously,another group cell transfected with empty vector was used as a control group. Results Compared with the control group, the expression of APOBEC3G was obviously increased in the HepG2. 2. 15 cell line transfected with pcDNA3.1 Human APOBEC3G-Myc-6Xllis at mRNA and protein levels. The amount of HBsAg and HBeAg in the supernatant of HepG2.2.15 cell line transfected with pcDNA3.1 Human APOBEC3G-Myc-6Xhis was apparently decreased by 70.38% and 62.88%,respectively. Conclusion It is powerfully verified that APOBEC3G can inhibit HBV replication in vitro. The result indicates APOBEC3G has potential to be chosed and developed as a novel anti-HBV agent.