title: Generation of a human hepatoma cell line supporting efficient replication of a lamivudine resistant hepatitis B virus.
abstract: Emergence of lamivudine (LAM) resistance causes treatment failure in patients with chronic hepatitis B and compromise the efficacy of subsequent salvage therapies with other nucleot(s)ide analogs (NAs). Establishment of cell-based assays supporting LAM-resistant hepatitis B virus (HBV) replication will not only provide tools for investigating the replication property, but also screening for antiviral agents efficiently inhibiting the replication of LAM-resistant HBV variants. Accordingly, a human hepatoma (HepG2)-derived cell line was established by stable transfection of a plasmid containing a 1.2 unit length of HBV genome harboring rtL180M and rtM204V mutations that confer LAM resistance. In addition to support efficient viral genome replication, the cell line also produces high levels of HBV virions and subviral particles. As expected, HBV DNA replication in this cell line is completely resistant to lamivudine, but sensitive to adefovir (ADV), entecavir (ETV) and tenofovir (TDF). The cell line is suitable for screening for antiviral agents that inhibit LAM-resistant HBV replication and inhibitors of HBsAg biosynthesis and secretion, which may reduce HBsAg antigenemia and ultimately help to restore host antiviral immune response against HBV and cure chronic HBV infection.
authors.affiliation: Key Laboratory of Medical Molecular Virology of the Ministries of Education and Health and Department of Infectious Disease, Huashan Hospital, Fudan University, Shanghai 200040, China.
authors.forename: Yijun
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Medical Molecular Virology of the Ministries of Education and Health and Department of Infectious Disease, Huashan Hospital, Fudan University, Shanghai 200040, China.
authors.forename: Yongmei
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Medical Molecular Virology of the Ministries of Education and Health and Department of Infectious Disease, Huashan Hospital, Fudan University, Shanghai 200040, China.
authors.forename: Yaoyue
authors.initials: Y
authors.lastname: Kang
authors.affiliation: Key Laboratory of Medical Molecular Virology of the Ministries of Education and Health and Department of Infectious Disease, Huashan Hospital, Fudan University, Shanghai 200040, China.
authors.forename: Jinyu
authors.initials: J
authors.lastname: Wang
authors.affiliation: Key Laboratory of Medical Molecular Virology of the Ministries of Education and Health and Department of Infectious Disease, Huashan Hospital, Fudan University, Shanghai 200040, China.
authors.forename: Hongyan
authors.initials: H
authors.lastname: Liu
authors.affiliation: Key Laboratory of Medical Molecular Virology of the Ministries of Education and Health and Department of Infectious Disease, Huashan Hospital, Fudan University, Shanghai 200040, China.
authors.forename: Haoxiang
authors.initials: H
authors.lastname: Zhu
authors.affiliation: Key Laboratory of Medical Molecular Virology of the Ministries of Education and Health and Department of Infectious Disease, Huashan Hospital, Fudan University, Shanghai 200040, China.
authors.forename: Yanli
authors.initials: Y
authors.lastname: Qin
authors.affiliation: Key Laboratory of Medical Molecular Virology of the Ministries of Education and Health and Department of Infectious Disease, Huashan Hospital, Fudan University, Shanghai 200040, China.
authors.forename: Richeng
authors.initials: R
authors.lastname: Mao
authors.affiliation: Key Laboratory of Infection and Oncology, Research Center of Molecular Medicine, Fujian Medical University, Fuzhou, China.
authors.forename: Xu
authors.initials: X
authors.lastname: Lin
authors.affiliation: Institute of Virology, University Hospital of Essen, Essen, Germany.
authors.forename: Mengji
authors.initials: M
authors.lastname: Lu
authors.affiliation: Key Laboratory of Medical Molecular Virology of the Ministries of Education and Health and Department of Infectious Disease, Huashan Hospital, Fudan University, Shanghai 200040, China. Electronic address: jmzhang@fudan.edu.cn.
authors.forename: Jiming
authors.initials: J
authors.lastname: Zhang
mesh_terms: D000225:Adenine / Q000031:analogs & derivatives / Q000494:pharmacology; D000998:Antiviral Agents / Q000494:pharmacology*; D024882:Drug Resistance, Viral; D006147:Guanine / Q000031:analogs & derivatives / Q000494:pharmacology; D056945:Hep G2 Cells; D006515:Hepatitis B virus / Q000187:drug effects* / Q000254:growth & development* / Q000502:physiology; D022781:Hepatocytes / Q000821:virology*; D006801:Humans; D019259:Lamivudine / Q000494:pharmacology*; D063065:Organophosphonates / Q000494:pharmacology; D000068698:Tenofovir; D014779:Virus Replication / Q000187:drug effects
chemical_list: D000998:Antiviral Agents; D063065:Organophosphonates; D019259:Lamivudine; C413685:entecavir; D006147:Guanine; C053001:adefovir; D000068698:Tenofovir; D000225:Adenine
keywords: HBsAg; HbeAg; Hepatitis B virus; Lamivudine resistance; Stable cell line