Aims:To examine the regulation of SREBP-1c and CAV1 by microRNA-29a (miR-29a) in cells infected with hepatitis C virus (HCV) in an attempt to control HCV-induced nonalcoholic fatty liver disease.Methods:In order to ex...Aims:To examine the regulation of SREBP-1c and CAV1 by microRNA-29a (miR-29a) in cells infected with hepatitis C virus (HCV) in an attempt to control HCV-induced nonalcoholic fatty liver disease.Methods:In order to examine the manipulation of SREBP-1c and CAV1 by miR-29a,oleic acid (OA)-treated JFH-I-infected Huh-7 cells were used.OA was added 24 h post-transfection and gene expression was investigated by qRT-PCR at 48 h post treatment.The functional impact of the observed alteration in SREBP-1c and CAV1 expression was analyzed by examining lipid droplet (LD) and triglyceride (TG) content at 72 h post-OA treatment using light microscopy and spectrophotometry,respectively.Viral load was quantified by qRT-PCR at 72 h post-transfection.Results:OA treatment induced the expression of miR-29a and SREBP-1c,as compared to untreated cells.Forced miR-29a expression led to a significant up-regulation of SREBP-1c as well as CAV1 compared to mock untransfected cells.Ectopic expression of miR-29a resulted in a marked increase in LDs and their respective TGs,while miR-29a antagomirs decreased both the LD and TG content compared to mock untransfected cells.Moreover,forcing the expression of miR-29a in JFH-1 HCV-infected Huh-7 cells resulted in 53% reduction in viral titers compared to mock untransfected Huh-7 cells.Conclusion:Inducing miR-29a expression significantly induces SREBP-1c and CAV1 expression,thereby increasing LDs as well as their respective TGs.Nonetheless,forcing the expression of miR-29a resulted in reduction of HCV RNA levels in Huh-7 cells.展开更多
文摘Aims:To examine the regulation of SREBP-1c and CAV1 by microRNA-29a (miR-29a) in cells infected with hepatitis C virus (HCV) in an attempt to control HCV-induced nonalcoholic fatty liver disease.Methods:In order to examine the manipulation of SREBP-1c and CAV1 by miR-29a,oleic acid (OA)-treated JFH-I-infected Huh-7 cells were used.OA was added 24 h post-transfection and gene expression was investigated by qRT-PCR at 48 h post treatment.The functional impact of the observed alteration in SREBP-1c and CAV1 expression was analyzed by examining lipid droplet (LD) and triglyceride (TG) content at 72 h post-OA treatment using light microscopy and spectrophotometry,respectively.Viral load was quantified by qRT-PCR at 72 h post-transfection.Results:OA treatment induced the expression of miR-29a and SREBP-1c,as compared to untreated cells.Forced miR-29a expression led to a significant up-regulation of SREBP-1c as well as CAV1 compared to mock untransfected cells.Ectopic expression of miR-29a resulted in a marked increase in LDs and their respective TGs,while miR-29a antagomirs decreased both the LD and TG content compared to mock untransfected cells.Moreover,forcing the expression of miR-29a in JFH-1 HCV-infected Huh-7 cells resulted in 53% reduction in viral titers compared to mock untransfected Huh-7 cells.Conclusion:Inducing miR-29a expression significantly induces SREBP-1c and CAV1 expression,thereby increasing LDs as well as their respective TGs.Nonetheless,forcing the expression of miR-29a resulted in reduction of HCV RNA levels in Huh-7 cells.
