title: The possible role of NS3 protease activity of hepatitis C virus on fibrogenesis and miR-122 expression in hepatic stellate cells.
abstract: The various roles of hepatitis C virus (HCV) NS3 protein in viral pathogenesis are emphasized, especially in the progression of fibrosis and tumors. The levels of miR-122 have been widely accepted as a critical factor in viral pathogenesis and disease progression. However, the possible correlation between miR-122 levels and fibrosis state has been less investigated. Therefore, in this study, plasmids expressing protease competent and protease mutated non-structural proteins 3 (NS3) were transfected into LX-2 cell line. Subsequently, the total RNA was extracted and real-time PCR was performed to measure the expression level of miR-122, collagen type 1 alpha 1 (COL1A1), alpha smooth muscle actin (α-SMA), and tissue inhibitor of metaloproteinase 1 (TIMP-1). Moreover, the transforming growth factor beta (TGF-β) levels in the supernatants of transfected cells were evaluated by ELISA. The gene expression analysis of fibrotic genes and TGF-β cytokine in LX-2 cells showed that protease competent NS3 had a significant fibrogenic impact when compared to protease defective NS3 or GFP control plasmids (P <0.001). The results also demonstrated that the expression of miR-122 was downregulated in both versions of the cells transfected with NS3 plasmids (P <0.01) irrespective of protease function. These results suggested that the protease function of NS3 protein is a crucial factor for the induction of hepatic fibrosis but it doesn't play a complete role in the expression of miR-122.
authors.forename: S
authors.initials: S
authors.lastname: Khanizadeh
authors.forename: M
authors.initials: M
authors.lastname: Ravanshad
authors.forename: S Y
authors.initials: SY
authors.lastname: Hosseini
authors.forename: P
authors.initials: P
authors.lastname: Davoodian
authors.forename: A N
authors.initials: AN
authors.lastname: Zadeh
authors.forename: F
authors.initials: F
authors.lastname: Sabahi
authors.forename: J
authors.initials: J
authors.lastname: Sarvari
authors.forename: Z
authors.initials: Z
authors.lastname: Khanlari
authors.forename: M
authors.initials: M
authors.lastname: Hasani-Azad
mesh_terms: D002460:Cell Line; D005337:Fibrin / Q000096:biosynthesis*; D005786:Gene Expression Regulation; D016174:Hepacivirus / Q000378:metabolism*; D055166:Hepatic Stellate Cells / Q000378:metabolism / Q000821:virology*; D006801:Humans; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D016212:Transforming Growth Factor beta / Q000235:genetics / Q000378:metabolism; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000378:metabolism*; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
chemical_list: D005337:Fibrin; D035683:MicroRNAs; D016212:Transforming Growth Factor beta; D017361:Viral Nonstructural Proteins; C531907:MIRN122 microRNA, human; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
keywords: HCV; miRNA; fibrosis; HSCs; NS3; protease; miR-122.