Effect And Clinical Significance Of MiRNA-548ah In Regulating The Replication And Expression Of Hepatitis B Virus Through Targeted HDAC4

Objective: 1.To investigate the effect and clinical significance of miRNA-548 ah targeted histone deacetylase 4(HDAC4)in regulating hepatitis B virus replication;2.To explore the expression and correlation of miRNA-548 ah and HDAC4 in liver tumor cell lines containing HBV and liver tissue from HBV infected patients.Methods: Expressions of miRNA-548 ah and HDAC4 in Huh7 cells,HepG2 cells,HepG2.2.15 cells and liver tissue of chronic hepatitis B patients were detected by fluorescent quantitative polymerase chain reaction.Huh7,HepG2 transfected by pHBV1.3 plasmid and HepG2.2.15 cells were transfected by mimics and inhibitors.The levels of hepatitis B e antigen(HBeAg),hepatitis B surface antigen(HBsAg)and HBV DNA in the supernatant were detected.Pearson Correlation Analysis was used for the expression of miRNA-548 ah and HDAC4 and clinical indexes in liver tissue of chronic hepatitis B patients.The t-test was used for comparison between two groups.SNK-q tests were used for multiple groups comparisons.Results: The expression of miRNA-548 ah and HDAC4 in Huh7 cells,HepG2 cells and HepG2.2.15 cells were significantly different(F=6.14,P=0.035;F=136.96,P=0.000).In HepG2 cells transfected with pHBV1.3 plasmid,the expression of HBsAg,HBeAg and HBV DNA were significantly increased in the miRNA-548 ah mimics group compared with the control group.The expressions of HBsAg,HBeAg and HBV DNA in the miRNA-548 ah inhibitor group were significantly decreased(F=20.61,276.87,110.03,P=0.000).In Huh7 cells transfected with pHBV1.3 plasmid,the expression of HBsAg,HBeAg and HBV DNA were significantly increased in the miRNA-548 ah mimics group compared with the control group.The expressions of HBsAg,HBeAg and HBV DNA were significantly reduced in the miRNA-548 ah inhibitor group(F=51.45,197.65,275.09,P=0.000).In HepG2.2.15 cells transfected with pHBV1.3 plasmid,the expression of HBsAg,HBeAg and HBV DNA were significantly increased in the miRNA-548 ah mimics group compared with the control group.The expressions of HBsAg,HBeAg and HBV DNA were significantly reduced in the miRNA-548 ah inhibitor group(F=73.09,163.48,3079.39,P=0.000).The expression of HDAC4 mRNA was increased in Huh7,HepG2 and HepG2.2.15 cells transfected with miRNA-548 ah inhibitor.However,the expression of HDAC4 mRNA was reduced in miRNA-548 ah mimics group.The expression of miRNA-548 ah in liver tissues of patients with chronic hepatitis B was significantly lower than that of patients with hepatitis B virus related cirrhosis(t=2.41,P=0.028).There was a significant negative correlation between the expression of miRNA-548 ah and HDAC4 in the liver tissues of patients with chronic hepatitis B(r=-0.746,P=0.021).The expression of HDAC4 in liver tissue was positively correlated with the level of serum ALT(r = 0.733,P = 0.025).Conclusions: 1.miRNA-548 ah could improve the replication and expression of HBV in the liver tumor cell lines containing HBV.2.HDACA4 may be the targeted gene regulated by miRNA-548 ah.3.miRNA-548 ah may be the marker of progression of CHB.4.HDAC4 may be related with the liver inflammatory activity of patients with CHB.