| Objective: This study aims to investigate that the interaction of HBV Precore 25 or Precore 22(P25,P22)and apolipoprotein B mRNA editing enzyme catalytic polypeptide-like 3A(APOBEC3A)in cells and its effect on HBV replication.Methods:Constructed P25 and P22 expression vectors,then,singleor co-transfected APOBEC3 A expression vector were transfected into HEK 293 T or Hep G2 cells.Western Blotting detected the expression of P25 and P22 and APOBEC3 A.Co-localization and interaction of APOBEC3 A and P25 or P22 were detected by confocal microscopy and co-immunoprecipitation respectively.Established loss of Heptitis B e antigen(HBeAg)expression cell model,and then,heptitis B surface antigen(HBsAg)and HBeAg in HepG2 culture supernatants were measured by enzyme-linked immunosorbent assay(ELISA),and,HBV DNA from core particles in transfected Hep G2 cells was detected in Southern blotting.Results:⑴Detected the expression of P25 and P22 in transfected HEK 293 T or HepG2 cells by Western Blotting.And its indicates that P25 and P22 expression vectors are constructed successfully.⑵Confocal microscopy and co-immunoprecipitation test that there have co-localization and interaction between APOBEC3 A and P25 or P22.⑶In loss of HBeAg expression cell model,the levels of HBsAg and HBeAg and HBV DNA from core particles are further reduced in co-transfected HepG2 cells,compared with those levels in HepG2 cells transfected with P25 or P22 or APOBEC3 A expression vectors alone.Conclusions: Our study confirms that P25,P22 and APOBEC3 A proteins can interact with each other and this interaction affects the replication of HBV.And,this conclusion supports the follow-up research “ HBV intracellular Precore protein may regulate IFN-α response through interacting with APOBEC3 A,which may be an important mechanism on the regulation of HBV replication and poorinterferon response”.Meanwhile,It also provides a basis for the research and development of new anti-hbv drugs in poor response of interferon treatment. |
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