APOBEC3F剪接亚型与全长APOBEC3F、APOBEC3G对乙肝病毒作用的比较

目的比较APOBEC3F剪接亚型3F79与完整APOBEC3F以及APOBEC3G对HepG2.2.15细胞中HBV DNA复制及HBsAg、HBeAg抗原分泌的影响。方法用PCR合成法扩增人APOBEC3F剪接亚型3F79,构建真核表达载体pEGFPC1-3F79和原核表达质粒pET28a-3F79,将pEGFPC1-3F79与含有全长APOBEC3F和APOBEC3G的质粒Pflag-APOBEC3F、PC-APOBEC3G-HA转染入HepG2.2.15中,检测转染后细胞上清中HBV DNA以及HBsAg和HBeAg的水平。结果构建的重组载体经酶切和PCR鉴定,表明3F79基因正确地插入。将pEGFPC1-3F79与Pflag-APOBEC3F、PC-APOBEC3G-HA转染HepG2.2.15细胞后,pEGFPC1-3F79对细胞中HBV的复制及HBsAg和HBeAg的分泌无明显的抑制作用(P0.05),而转染含有完整APOBEC3F和APOBEC3G质粒的HepG2.2.15细胞与对照组相比,HBsAg、HBeAg、HBV DNA含量明显下降,差异有统计学意义(P0.05)。结论 3F79不能像完整APOBEC3F和APOBEC3G一样抑制HepG2.2.15细胞中HBV DNA复制以及抗原的分泌。

Comparison of truncated APOBEC3F with complete APOBEC3F and APOBEC3G on HBV

Objective To evaluate the effect of truncated APOBEC3F（3F79 ） on hepatitis B virus in HepG2.2.15 cells compared with complete APOBEC3F and APOBEC3G. Meth The gene of truncated APOBEC3F was obtained with PCR synthetic method and then cloned in eukaryotic expression vector pEGFP-C1 and prokaryotic expression vector pET28a to construct pEGFPC1-3F79 and pET28a-3F79, pEGFPC1-3F79 as well as Pflag-APOBEC3F and PC-APO- BEC3G-HA was transfected in HepG2.2. 15 cells, respectively. Then the concentrations of HBV DNA, HBsAg and HBeAg in each group were quantified. Results Restriction enzyme and PCR analysis indicated that the 3F79 gene was correctly inserted into pEGFP-C1 and pET28a. 3F79, complete APOBEC3F and APOBEC3G were expressed in cytoplasm by fluorescence microscope after pEGFPC1-3F79 as well as Pflag-APOBEC3F and PC-APOBEC3G-HA were transfected in HepG2. pEGFPC1-3F79 had no significant effects on HBV replication in HepG2.2.15 cells, while the transfection of vectors containing complete APOBEC3F and APOBEC3G significantly decreased the levels of HBV DNA, HBsAg and HBeAg, respectively （P 〈 0.05）. Conclusion 3F79 do not have similar effects on HBV DNA replication and HBsAg and HBeAg secretion after being transfected into HcpG2.2. 15 as vectors containing complete APOBEC3F and APO- BEC3G.