title: MicroRNA-34a Promotes Hepatic Stellate Cell Activation via Targeting ACSL1.
abstract: BACKGROUND
  The incidence of liver fibrosis remains high due to the lack of effective therapies. Our previous work found that microRNA (miR)-34a expression was increased, while acy1-CoA synthetase long-chain family member1 (ACSL1) was decreased, in a dimethylnitrosamine (DNS)-induced hepatic fibrosis rat model. We hypothesized that miR-34a may play a role in the process of hepatic fibrosis by targeting ACSL1.
  
  
  METHODS
  From days 2 to 14, cultured primary hepatic stellate cells (HSCs) underwent cell morphology, immunocytochemical staining, and quantitative reverse transcription PCR (RT-qPCR) for alpha smooth muscle actin (a-SMA), desmin, rno-miR-34a, and ACSL1 expression. Wild-type and mutant luciferase reporter plasmids were constructed according to the predicted miR-34a binding site on the 3'-untranslated region (UTR) of the ACSL1 mRNA and then transfected into HEK293 cells. rno-miR-34a was silenced in HSCs to confirm that rno-miR-34a negatively regulates ACSL1 expression. mRNA and protein expression of α-SMA, type I collagen, and desmin were assayed in miR-34a-silenced HSCs.
  
  
  RESULTS
  HSCs were deemed quiescent during the first 3 days and activated after 10 days. rno-miR-34a expression increased, and ACSL1 expression decreased, from day 2 to 7 to 14. rno-miR-34a was shown to specifically bind to the 3'-UTR of ACSL1. miR-34a-silenced HSCs showed higher ACSL1and lower α-SMA, type I collagen, and desmin expression than that of matching negative controls and non-transfected cells.
  
  
  CONCLUSIONS
  miR-34a appears to play an important role in the process of liver fibrosis by targeting ACSL1 and may show promise as a therapeutic molecular target for hepatic fibrosis.
authors.affiliation: Department of Pathology, Changzheng Hospital, The Second Military Medical University, Shanghai, China (mainland).
authors.forename: Gangli
authors.initials: G
authors.lastname: Yan
authors.affiliation: Department of Pathology, Changzheng Hospital, The Second Military Medical University, Shanghai, China (mainland).
authors.forename: Binbin
authors.initials: B
authors.lastname: Li
authors.affiliation: Department of Pathology, Changzheng Hospital, The Second Military Medical University, Shanghai, China (mainland).
authors.forename: Xuan
authors.initials: X
authors.lastname: Xin
authors.affiliation: Department of Pathology, Changzheng Hospital,the Second Military Medical University, Shanghai, China (mainland).
authors.forename: Midie
authors.initials: M
authors.lastname: Xu
authors.affiliation: State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai, China (mainland).
authors.forename: Guoqing
authors.initials: G
authors.lastname: Ji
authors.affiliation: Department of Pathology, Changzheng Hospital, The Second Military Medical University, Shanghai, China (mainland).
authors.forename: Hongyu
authors.initials: H
authors.lastname: Yu
mesh_terms: D020413:3' Untranslated Regions; D000199:Actins / Q000378:metabolism; D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000378:metabolism*; D004128:Dimethylnitrosamine; D005786:Gene Expression Regulation; D020868:Gene Silencing; D057809:HEK293 Cells; D055166:Hepatic Stellate Cells / Q000166:cytology*; D006801:Humans; D007150:Immunohistochemistry; D008099:Liver / Q000187:drug effects / Q000473:pathology; D008103:Liver Cirrhosis / Q000473:pathology; D008297:Male; D035683:MicroRNAs / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D020413:3' Untranslated Regions; D000199:Actins; D051792:Basic Helix-Loop-Helix Proteins; D004128:Dimethylnitrosamine; D035683:MicroRNAs; C065500:ASCL1 protein, human; C499318:Ascl1 protein, rat; C522425:MIRN34 microRNA, human; C560944:MIRN34 microRNA, rat