MicroRNA-196 represses Bach1 protein and hepatitis C virus gene expression in human hepatoma cells expressing hepatitis C viral proteins
Open Access
- 23 October 2009
- journal article
- viral hepatitis
- Published by Ovid Technologies (Wolters Kluwer Health) in Journal of Hepatology
- Vol. 51 (5), 1494-1504
- https://doi.org/10.1002/hep.23401
Abstract
Hepatitis C virus (HCV) directly induces oxidative stress and liver injury. Bach1, a basic leucine zipper mammalian transcriptional repressor, negatively regulates heme oxygenase 1 (HMOX1), a key cytoprotective enzyme that has antioxidant and anti‐inflammatory activities. microRNAs (miRNAs) are small noncoding RNAs (≈22 nt) that are important regulators of gene expression. Whether and how miRNAs regulate Bach1 or HCV are largely unknown. The aims of this study were to determine whether miR‐196 regulates Bach1, HMOX1, and/or HCV gene expression. HCV replicon cell lines (Con1 and 9–13) of the Con1 isolate and J6/JFH1‐based HCV cell culture system were used in this study. The effects of miR‐196 mimic on Bach1, HMOX1, and HCV RNA, and protein levels were measured by way of quantitative real‐time polymerase chain reaction (qRT‐PCR) and Western blotting, respectively. The Dual Glo Luciferase Assay System was used to determine reporter activities. miR‐196 mimic significantly down‐regulated Bach1 and up‐regulated HMOX1 gene expression and inhibited HCV expression. Dual luciferase reporter assays demonstrated that transfection of miR‐196 mimic resulted in a significant decrease in Bach1 3′‐untranslated region (UTR)–dependent luciferase activity but not in mutant Bach1 3′‐UTR–dependent luciferase activity. Moreover, there was no detectable effect of mutant miR‐196 on Bach1 3′‐UTR–dependent luciferase activity. Conclusion: miR‐196 directly acts on the 3′‐UTR of Bach1 messenger RNA and translationally represses the expression of this protein, and up‐regulates HMOX1. miR‐196 also inhibits HCV expression in HCV replicon cell lines (genotype 1b) and in J6/JFH1 (genotype 2a) HCV cell culture system. Thus, miR‐196 plays a role in both HMOX1/Bach1 expression and the regulation of HCV expression in human hepatocytes. Overexpression of miR‐196 holds promise as a potential novel strategy to prevent or ameliorate hepatitis C infection, and to protect against liver injury in chronic HCV infection. (HEPATOLOGY 2010.)Keywords
This publication has 46 references indexed in Scilit:
- Position-Dependent Function for a Tandem MicroRNA miR-122-Binding Site Located in the Hepatitis C Virus RNA GenomeCell Host & Microbe, 2008
- Heme oxygenase-1 suppresses hepatitis C virus replication and increases resistance of hepatocytes to oxidant injuryJournal of Hepatology, 2008
- Zinc mesoporphyrin induces rapid and marked degradation of the transcription factor bach1 and up-regulates HO-1Biochimica et Biophysica Acta (BBA) - Gene Regulatory Mechanisms, 2008
- A viral microRNA functions as an orthologue of cellular miR-155Nature, 2007
- Kaposi's Sarcoma-Associated Herpesvirus Encodes an Ortholog of miR-155Journal of Virology, 2007
- Ultra-Rapid Cardiotoxicity of the Hepatitis C Virus Protease Inhibitor BILN 2061 in the Urokinase-Type Plasminogen Activator MouseGastroenterology, 2007
- Reciprocal Effects of Micro-RNA-122 on Expression of Heme Oxygenase-1 and Hepatitis C Virus Genes in Human HepatocytesGastroenterology, 2007
- Interferon modulation of cellular microRNAs as an antiviral mechanismNature, 2007
- Conserved Seed Pairing, Often Flanked by Adenosines, Indicates that Thousands of Human Genes are MicroRNA TargetsCell, 2005
- The functions of animal microRNAsNature, 2004