O-糖基转移酶、Glt25D2与乙型肝炎包膜蛋白大蛋白的分泌有关

Glt25D2, an O-glycosyltransferase is related with the secretion of hepatitis B virus surface large protein

目的在前期研究结果提示Glt25D2与HBV包膜蛋白大蛋白(LHBs)在体外相互作用基础上证明Glt25D2是否与HBV LHBs分泌相关。方法采用激光共聚焦方法分析Glt25D2与LHBs在HepG2细胞内的定位。免疫共沉淀方法进一步证实Glt25D2与LHBs的相互作用。采用实时荧光定量PCR和Western blot方法分析mRNA和蛋白表达水平。应用ELISA方法检测细胞上清LHBs水平。实时荧光定量PCR方法检测上清HBV病毒载量。ELISA方法检测上清HBV LHBs水平。Western blot方法检测细胞中LHBs蛋白含量。Cobas Amplicor HBV Monitor Test方法检测细胞上清液HBV DNA载量。结果 Glt25D2与LHBs在体外相互作用,上调的Glt25D2高表达促进HBV DNA复制和LHBs表达,Glt25D2低表达抑制HBV DNA复制和LHBs表达。结论 Glt25D2与乙型肝炎包膜蛋白大蛋白的分泌有关。

Objective Our previous study demonstrated that Glt25D2 interacts with HBV LHBs in vitro. In this study, we aim to prove if Glt25D2, an O-glycosyltransferase, is related with the secretion of hepatitis B virus surface large protein （LHBs）. Methods Confocal microscopy was used to determine the co-location between Glt25D2 and LHBs in HepG2 cells. The interaction between Glt25D2 and LHBs in vitro was examined by the method of co-immunoprecipitation. Quantitative real-time PCR and Western blot were undertaken to evaluate the levels of mRNA and protein. The levels of LHBs in the supernatant were measured with ELISA kits and HBV DNA was quantified with the Cobas Amplicor HBV Monitor Test. Results Our data demonstrated that Glt25D2 interactes with LHBs in vitro, up-regulated Glt25D2 expression could increase HBV DNA and LHBs levels in HepG2.2.15, down-regulated Glt25D2 expression decreased HBV DNA and LHBs levels. Conclusions Our results suggest that Glt25D2 is related with HBV LHBs secretion.