The inhibition effect of GLS4JHS on the transcription activity of covalently closed circular DNA in HepAD38 cells / 中华传染病杂志

ABSTRACT

Objective The hepatitis B virus (HBV) core protein assembly inhibitors GLS4JHS could destroy HBV capsid assembly and the formation of non-capsid polymer structure.The aim of this study is to explore the mechanisms of GLS4JHS in inhibiting HBV replication.Methods HepAD38 cells was used as the study model.TaqMan real-time polymerase chain reaction (PCR) and quantitative real-time PCR with specific primers were used to measure the change in pregenomic RNA (pgRNA) and covalently closed circular DNA (cccDNA) levels under different concentrations.ChIP assay in HepAD38 cells was used to assess the recruitment of HBV core protein and histone modifications.Results The amount of cccDNA and pgRNA decreased with the increasing GLS4JHS concentrations.After the drug concentrations reached 400 nmol/L, cccDNA and pgRNA declined by 94% and 84%, respectively.Both HBV core protein occupancy on the cccDNA and cccDNA-bound H3 histone acetylation were reduced by GLS4JHS.Conclusions GLS4JHS decreases transcriptional activity of cccDNA and reduces pgRNA production by inhibiting cccDNA minichromosome bound to HBV core protein and acetylated histone H3, which results in HBV DNA formation.