Original Article

Archives of Virology

, Volume 154, Issue 7, pp 1021-1026

First online:

Establishment of cell lines using a doxycycline-inducible gene expression system to regulate expression of hepatitis B virus X protein

  • Hong TangAffiliated withCenter of Infectious Diseases, Division of Molecular Biology of Infectious Diseases, National Key Laboratory of Biotherapy (Sichuan University), West China Hospital of Sichuan University
  • , Li LiuAffiliated withCenter of Infectious Diseases, Division of Molecular Biology of Infectious Diseases, National Key Laboratory of Biotherapy (Sichuan University), West China Hospital of Sichuan University
  • , Feng-Jun LiuAffiliated withCenter of Infectious Diseases, Division of Molecular Biology of Infectious Diseases, National Key Laboratory of Biotherapy (Sichuan University), West China Hospital of Sichuan University
  • , En-Qiang ChenAffiliated withCenter of Infectious Diseases, Division of Molecular Biology of Infectious Diseases, National Key Laboratory of Biotherapy (Sichuan University), West China Hospital of Sichuan University
  • , Seishi MurakamiAffiliated withDepartment of Molecular Oncology, Cancer Research Institute, Kanazawa University
  • , Yong LinAffiliated withCenter of Infectious Diseases, Division of Molecular Biology of Infectious Diseases, National Key Laboratory of Biotherapy (Sichuan University), West China Hospital of Sichuan University
  • , Fang HeAffiliated withCenter of Infectious Diseases, Division of Molecular Biology of Infectious Diseases, National Key Laboratory of Biotherapy (Sichuan University), West China Hospital of Sichuan University
  • , Tao-You ZhouAffiliated withCenter of Infectious Diseases, Division of Molecular Biology of Infectious Diseases, National Key Laboratory of Biotherapy (Sichuan University), West China Hospital of Sichuan University
  • , Fei-Jun HuangAffiliated withDepartment of Forensic Pathology, College of Basic Medicine and Forensic Medicine, Sichuan University Email author 

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Abstract

The hepatitis B virus (HBV) X gene plays an important role in HBV-associated pathogenesis, especially hepatocarcinogenesis. Establishment of a stable and regulable HBx expression system will allow study of the function of this gene. Here, we describe the development of a doxycycline-inducible recombinant plasmid (pBPSTR3-FlagX) with the full-length HBV X gene and all components of the tetracycline-on (“Tet-on”) gene expression system. This vector exhibited dose-dependent doxycycline-dependent induction of the Flag-HBx protein in HepG2 and Hep3B cells. We also observed dose-dependent doxycycline transactivation of HBx in HepG2 cells. After transfecting HepG2 cells with the pBPSTR3-FlagX plasmid, we isolated five puromycin-resistant cell clones with stable HBx expression, two of which exhibited stable and tight control of HBx expression by doxycycline. This new system has great potential for functional studies of the HBV X gene.