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06 October 2015 : Laboratory Research  

MicroRNA-34a Promotes Hepatic Stellate Cell Activation via Targeting ACSL1

Gangli YanBE, Binbin LiCD, Xuan XinDF, Midie XuDF, Guoqing JiCF, Hongyu YuAEG

DOI: 10.12659/MSM.894000

Med Sci Monit 2015; 21:3008-3015

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.

MATERIAL AND 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.

Keywords: 3' Untranslated Regions, Basic Helix-Loop-Helix Transcription Factors - metabolism, Dimethylnitrosamine, Gene Expression Regulation, Gene Silencing, HEK293 Cells, Hepatic Stellate Cells - cytology, Liver - pathology, Liver Cirrhosis - pathology

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750