Study On The Effect Of PA On The Expression Of HNF3α/3β/4α In Vitro

A lot of liver enriched transcriptional factors,including HNF3a,HNF3β,HNF4a,form a very complicated regulation network.They not only regulate the expression of specified gene and differentiation of cell in liver,but also affect the life cycle of HBV in liver.HNF4a,which is a central regulator,can play the role of liver enriched transcriptional factors,It also regulate the expression of other hepatocyte nuclear factors such as HNF3.In HepG2.215 cell,PA is able to inhibit the secretion of HBV antigen and DNA replication.3TC and PA combination restrain HBx into cellular nucleus that is very important to HBV DNA replication in liver.And the inhibitory effect of their combination was more effective than that of 3TC.This paper investigated the influence of PA on HNF in Huh7 and HepG2.215 cells.The experimental result showed that:(1)PA(10μg/ml)had no effect of the cell proliferation and had no toxic effect on Huh7 cells.PA(10μg/ml)not only had no toxic effect on HepG2.215 cells,but also promote proliferation of HepG2.215 cells.And the promotion effect is dependent on time.(2)HepG2.215 cells are used in subsequent experiment after G418 screening that all cells can stably express HBV antigen.(3)PA(10μg/ml)can not only inhibit the mRNA expression of HNF3α/HNF3β/HNF4a but also the protein expression of HNF4a in Huh7 cells.(4)PA(10γμg/ml)can not only inhibit the mRNA expression of HNF4a but also the protein expression of HNF4a in HepG2.215 cells.Above all,the research of this paper showed that PA can inhibit the mRNA expression of HNF3a/HNF3β/HNF4α in Huh7 cells,we can speculate that PA may influence the expression of specified gene and differentiation of cell in liver by acting on different hepatocyte nuclear factors,but the specific therapeutic effects need further experiments to explore.PA can inhibit both mRNA and protein expression of HNF4a in HepG2.215 cells where HBV DNA can replicate.We can infer that PA may influence the level of replication of HBV DNA in HepG2.215 cells directly or indirectly by reducing HNF4a,but this speculation still needs more experiments to verify.On the other hand,It can not explain why 3TC and PA combination restrain HBx into cellular nucleus.In a word,It is very necessary to do more experiments to clarify the molecular mechanism of PA against HBV.