title: MicroRNA-122-dependent and -independent replication of Hepatitis C Virus in Hep3B human hepatoma cells.
abstract: The study of Hepatitis C Virus (HCV) has benefitted from the use of the Huh7 cell culture system, but until recently there were no other widely used alternatives to this cell line. Here we render another human hepatoma cell line, Hep3B, permissive to the complete virus life cycle by supplementation with the liver-specific microRNA miR-122, known to aid HCV RNA accumulation. When supplemented, Hep3B cells produce J6/JFH-1 virus titres indistinguishable from those produced by Huh7.5 cells. Interestingly, we were able to detect and characterize miR-122-independent replication of di-cistronic replicons in Hep3B cells. Further, we show that Argonaute-2 (Ago2) is required for miR-122-dependent replication, but dispensable for miR-122-independent replication, confirming Ago2's role in mediating the activity of miR-122. Thus Hep3B cells are a model system for the study of HCV, and miR-122 independent replication is a model to identify proteins involved in the function of miR-122.
authors.affiliation: Department of Microbiology and Immunology, Saskatoon, Canada SK S7N 5E5. patricia.thibault@usask.ca
authors.forename: Patricia A
authors.initials: PA
authors.lastname: Thibault
authors.forename: Adam
authors.initials: A
authors.lastname: Huys
authors.forename: Pearl
authors.initials: P
authors.lastname: Dhillon
authors.forename: Joyce A
authors.initials: JA
authors.lastname: Wilson
mesh_terms: D060565:Argonaute Proteins / Q000378:metabolism*; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000821:virology*; D045744:Cell Line, Tumor; D016174:Hepacivirus / Q000502:physiology*; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics / Q000821:virology; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D034741:RNA, Small Interfering; D012367:RNA, Viral / Q000378:metabolism; D019562:Viral Load; D014779:Virus Replication
chemical_list: C466422:AGO2 protein, human; D060565:Argonaute Proteins; C531907:MIRN122 microRNA, human; D035683:MicroRNAs; D034741:RNA, Small Interfering; D012367:RNA, Viral