Formation of covalently closed circular DNA in Hep38.7-Tet cells, a tetracycline inducible hepatitis B virus expression cell line

Biochem Biophys Res Commun. 2014 Sep 26;452(3):315-21. doi: 10.1016/j.bbrc.2014.08.029. Epub 2014 Aug 20.

Abstract

Hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) plays a central role in chronic HBV infection. However, analysis of the molecular mechanism of cccDNA formation is difficult because of the low efficiency in tissue cultured cells. In this study, we developed a more efficient cccDNA expression cell, Hep38.7-Tet, by subcloning from a tetracycline inducible HBV expression cell, HepAD38. Higher levels of cccDNA were produced in Hep38.7-Tet cells compared to HepAD38 cells. In Hep38.7-Tet cells, the cccDNA was detectable at six days after HBV induction. HBV e antigen (HBeAg) secretion was dependent upon cccDNA production. We screened chemical compounds using Hep38.7-Tet cells and HBeAg secretion as a marker. Most of the hit compounds have already been reported as anti-HBV compounds. These data suggested that Hep38.7-Tet cells will be powerful tools for analysis of the molecular mechanism of cccDNA formation/maintenance and development of novel therapeutic agents to control HBV infection.

Keywords: Antiviral; HBV; cccDNA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antiviral Agents / pharmacology
  • Cell Line, Transformed / drug effects*
  • Cell Line, Transformed / metabolism
  • Cell Line, Transformed / virology
  • DNA, Circular / genetics*
  • DNA, Circular / metabolism
  • DNA, Viral / genetics*
  • DNA, Viral / metabolism
  • Founder Effect
  • Gene Expression
  • Hepatitis B e Antigens / biosynthesis
  • Hepatitis B e Antigens / genetics
  • Hepatitis B virus / drug effects*
  • Hepatitis B virus / genetics
  • Hepatitis B virus / metabolism
  • Hepatocytes / drug effects*
  • Hepatocytes / metabolism
  • Hepatocytes / virology
  • High-Throughput Screening Assays
  • Humans
  • Protein Synthesis Inhibitors / pharmacology
  • Small Molecule Libraries / pharmacology
  • Tetracycline / pharmacology
  • Virus Replication / drug effects

Substances

  • Antiviral Agents
  • DNA, Circular
  • DNA, Viral
  • Hepatitis B e Antigens
  • Protein Synthesis Inhibitors
  • Small Molecule Libraries
  • Tetracycline