Effect And Mechnism Of MiRNA-146a On The Life Cycle Of Hepatitis B Virus

Objective: To investigate the effect of microRNA-146a(miR-146a)on the life cycle of hepatitis B virus(HBV)and mechanisms.Methods: The miRNA expression profile differences were compared by miRNA array between HepG2 and HepG2.2.15 cells and interested miR-146 a was confirmed by RT-PCR.MiR-146 a mimic and inhibitor were transfected into HepG2.2.15 cells respectively,followed by the quantification of HBV replication by RT-PCR,and protein expression levels by ELISA and western blot.Dual-luciferase reporter assay was used to detect the interaction between miR-146 a and potential target HS3ST3B1.Results:(1)Totally 74 miRNAs expression levels changed in HepG2.2.15 cells,including 29 up-regulated and 45 down-regulated.MiR-146 a level in HepG2.2.15 cells was significantly higher than that in HepG2 by RT-PCR(P<0.05).(2)When miR-146 a was transfected into HepG2.2.15 cells,the HBV replication and protein levels were significantly higher than control groups(P<0.05);Vice versa(P<0.05).(3)Bioinformatics analysis showed that HS3ST3B1 was a potential target of mi R-146 a.The reporter luciferase reporter system showed that the reported fluorescence of HS3ST3B1 wild type vector was significantly lower than that in the control group(P<0.05).There was no significant difference in the fluorescence value between HS3ST3B1 mutant vector and control group(P> 0.05).HepG2.2.15 cells were transfected with miR-146 a mimic,compared with the control group,the mRNA level of HS3ST3B1 in the experimental group was not significantly changed(P> 0.05),and the level of HS3ST3B1 protein in the experimental group was significantly decreased(P<0.05).Conclusions: This study confirmed that miR-146 a can affect the life cycle of HBV,further study of its mechanism found that mi R-146 a may affect HBV life cycle by acting on HBV inhibitory factor HS3ST3B1 3?UTR and inhibition of its translation.