title: miR-29a Promotes Lipid Droplet and Triglyceride Formation in HCV Infection by Inducing Expression of SREBP-1c and CAV1.
abstract: 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 non-alcoholic 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.
authors.affiliation: Department of Pharmaceutical Chemistry, German University in Cairo, New Cairo City, Egypt.
authors.forename: Mennatallah Mamdouh
authors.initials: MM
authors.lastname: Mahdy
authors.affiliation: Department of Pharmacology and Toxicology, German University in Cairo, New Cairo City, Egypt; School of Medicine, NewGiza University, Cairo, Egypt.
authors.forename: Nada Magdy
authors.initials: NM
authors.lastname: El-Ekiaby
authors.affiliation: Department of Pharmaceutical Biology, German University in Cairo, New Cairo City, Egypt.
authors.forename: Rana Mahmoud
authors.initials: RM
authors.lastname: Hashish
authors.affiliation: Department of Stem Cells and Regenerative Medicine, Zewail City of Science and Technology, Giza, Egypt.
authors.forename: Radwa Ayman
authors.initials: RA
authors.lastname: Salah
authors.affiliation: Department of Pharmaceutical Chemistry, German University in Cairo, New Cairo City, Egypt.
authors.forename: Rasha Sayed
authors.initials: RS
authors.lastname: Hanafi
authors.affiliation: Department of Chemistry, Novel Diagnostic & Therapeutics, American University in Cairo, New Cairo City, Egypt.
authors.forename: Hassan Mohamed
authors.initials: HM
authors.lastname: Azzazy
authors.affiliation: Department of Pharmacology and Toxicology, German University in Cairo, New Cairo City, Egypt; School of Medicine, NewGiza University, Cairo, Egypt.
authors.forename: Ahmed Ihab
authors.initials: AI
authors.lastname: Abdelaziz
keywords: Caveolin-1; HCV; Lipid droplets; MicroRNA-29a; SREBP-1c