miR-29a Promotes Lipid Droplet and Triglyceride Formation in HCV Infection by Inducing Expression of SREBP-1c and CAV1  

作　　者：Mennatallah Mamdouh Mahdy Nada Magdy El-Ekiaby Rana Mahmoud Hashish Radwa Ayman Salah Rasha Sayed Hanafi Hassan Mohamed El-Said Azzazy Ahmed Ihab Abdelaziz 

机构地区：Department of Pharmaceutical Chemistry, German University in Cairo, New Cairo City, Egypt Department of Pharmacology and Toxicology, German University in Cairo, New Cairo City, Egypt School of Medicine, New Giza University, Cairo, Egypt Department of Pharmaceutical Biology, German University in Cairo, New Cairo City, Egypt Department of Stem Cells and Regenerative Medicine, Zewail City of Science and Technology,Giza, Egypt Department of Chemistry, Novel Diagnostic & Therapeutics, American University in Cairo, New Cairo City, Egypt

出　　处：《Journal of Clinical and Translational Hepatology》2016年第4期293-299,共7页临床与转化肝病杂志（英文版）

摘　　要：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.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.

关 键 词：CAVEOLIN-1 HCV Lipid droplets MicroRNA-29a SREBP-1C 

分 类 号：R73[医药卫生—肿瘤]