Clinical–liver, pancreas, and biliary tractReciprocal Effects of Micro-RNA-122 on Expression of Heme Oxygenase-1 and Hepatitis C Virus Genes in Human Hepatocytes
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Materials
The human hepatoma cell line, Huh-7, was purchased from the Japan Health Research Resources Bank (Osaka, Japan). Ferric (Fe+3)-protoporphyrin IX•Cl (hemin) and cobalt protoporphyrin (CoPP) were from Porphyrin Products (Logan, UT). Goat anti-human glyceraldehyde phosphate dehydrogenase (GAPDH) antibody was purchased from Santa Cruz Biotechnology, Inc (Santa Cruz, CA). Mouse antihepatitis C core protein was purchased from Affinity BioReagents, Inc (Golden, CO). Mouse anti-HCV nonstructural
Down-Regulation of HCV RNA and Protein Expression by Antagomir of miR-122 in CNS3 and 9-13 Replicon Cells
miR-122 is specifically expressed and highly abundant in the human liver, in which it constitutes 70% of the total miRNA population.47, 50 There are 2 predicted binding sites for miR-122 in HCV viral 3′ and 5′ NCRs.53, 55 To assess down-regulation of antagomir of miR-122 on HCV expression, Huh-7 cells stably replicating HCV subgenomic (NS3-5B) replicon were used (Figure 1A). First, we compared the 2 cell lines stably replicating HCV core-NS3 or NS3-NS5B proteins with WT Huh-7 cells and cells
Discussion
The major findings of this study are that (1) an antagomir of miR-122 reduces the abundance of HCV mRNA and protein in CNS3 and 9-13 cells; (2) transfection with mimic-miR-122 increases HCV levels, whereas transfection with NSCA did not change the level of HCV, when compared with the cells that were not transfected; (3) antagomir of miR-122 decreased Bach1 and increased HO-1 mRNA levels in CNS3 and 9-13 replicon cells; (4) silencing Bach1 with Bach1-siRNA increases HO-1 mRNA levels and
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The opinions expressed herein are those of the authors. They do not necessarily reflect the official views of the U.S.P.H.S. or The University of Connecticut Health Center.
No conflicts of interest exist.
Supported by NIH RO1-DK38825 and contracts NO-1 DK92326 and UO-1 DK065193.