Characterization of novel human hepatoma cell lines with stable hepatitis B virus secretion for evaluating new compounds against lamivudine- and penciclovir-resistant virus.
L-Nucleoside analogs are new therapeutic agents for treatment of chronic hepatitis B. However, their clinical application was limited by the emergence of viral resistance. It is important to develop a new system to evaluate drug cross-resistance and to test new agents that may overcome resistant virus. In this report, three cell lines HepG2-WT10, HepG2-SM1, and HepG2-DM2 are presented; these cell lines were established by transfection of HepG2 cells with unique fully functional 1.1x hepatitis B virus (HBV) genomes: wild-type HBV-adr and its L526M and L526MM550V variants, respectively. We have demonstrated that these genomes have different susceptibilities to lamivudine [L(-)SddC] and penciclovir (PCV). By examining HBV RNA transcription, antigen expression, progeny DNA replication, and viral susceptibilities to L(-)SddC, PCV, and other nucleoside analogs, it is concluded that the cell lines are able to stably produce L(-)SddC- and PCV-sensitive and -resistant HBV virions. In addition, the relative susceptibilities of the wild-type and mutant HBV produced from the stably transfected cell lines to several anti-HBV nucleoside analogs were also examined and found to be about the same as those found by using a transient infection system. PMEA [9-(2-phosphonylmethoxytehyl)-adenine] and QYL685 are able to suppress L(-)SddC- and PCV-resistant HBV. In conclusion, this cell culture system is a novel and useful tool for evaluating anti-HBV compounds and biologics.
Fu
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Department of Pharmacology, Yale University School of Medicine, New Haven, Connecticut 06520, USA.
Cheng
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D000212:Acyclovir / Q000031:analogs & derivatives* / Q000494:pharmacology; D000956:Antigens, Viral / Q000378:metabolism; D000998:Antiviral Agents / Q000494:pharmacology*; D006528:Carcinoma, Hepatocellular / Q000821:virology; D004352:Drug Resistance, Microbial / Q000502:physiology; D006147:Guanine; D006515:Hepatitis B virus / Q000187:drug effects* / Q000235:genetics / Q000502:physiology; D006801:Humans; D019259:Lamivudine / Q000494:pharmacology; D008113:Liver Neoplasms / Q000821:virology; D008826:Microbial Sensitivity Tests / Q000379:methods*; D013997:Time Factors; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000821:virology; D014779:Virus Replication / Q000187:drug effects
D000956:Antigens, Viral; D000998:Antiviral Agents; D019259:Lamivudine; C053539:penciclovir; D006147:Guanine; D000212:Acyclovir