Screening and identification of compounds inhibiting transcriptional activity of HBV core promoter

Abstract

Objective To construct a stable cell line targeting the transcriptional activity of core promoter for screening drugs that affect the transcription of HBV. Methods HepG2-Pcore-Gluc was constructed by recombinant lentivirus to stably express Gaussia luciferase driven by HBV core promoter in HepG2 cells, and then the luciferase was detected to indicate the activity of the core promoter. Southern blotting, fluorescence quantitative PCR and other experiments were used to further evaluate the effects of the screened compounds on the levels of HBV RNA and HBV DNA after several rounds of screening in HBV-replication cell models. Results Forty-one compounds that could affect luciferase activity larger than 2 times were screened out of 1 450 compounds in the first round screening by using HepG2-Pcore-Gluc cell model, which were tested in a second round of dose-dependent test to produce 2 compounds: Reversine and SCH58261. Further, Reversine effectively decreased the levels of HBV RNA, HBV DNA, HBeAg and HBsAg in transiently-transfected HepG2 and HepG2.2.15 cells. Conclusion Reversine is identified by using a novel cell model (HepG2-Pcore-Gluc) which targets the transcription of HBV core promoter. These results provide evidences that Reversine acts through inhibiting the transcriptional activity of core promoter.

Keywords

• reversine
• hepititis b virus
• core promoter
• cell model
• compounds screening