Targeting EZH1 and EZH2 contributes to the suppression of fibrosis-associated genes by miR-214-3p in cardiac myofibroblasts

// Wen-Si Zhu 1,2,* , Chun-Mei Tang 2,3,* , Zhen Xiao 1,2,* , Jie-Ning Zhu 1,2 , Qiu-Xiong Lin 1,2 , Yong-Heng Fu 1,2 , Zhi-Qin Hu 2,3 , Zhuo Zhang 2,4 , Min Yang 1,2 , Xi-Long Zheng 5 , Shu-Lin Wu 1,2 and Zhi-Xin Shan 1,2 1 Guangdong Cardiovascular Institute, Guangdong Provincial Key Laboratory of Clinical Pharmacology, Guangzhou, China 2 Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangzhou, China 3 Southern Medical University, Guangzhou, China 4 School of Medicine, South China University of Technology, Guangzhou, China 5 The Libin Cardiovascular Institute of Alberta, Department of Biochemistry & Molecular Biology, The University of Calgary, Calgary, Canada * These authors have contributed equally to this work Correspondence to: Zhi-Xin Shan, email: // Keywords : microRNA-214-3p, cardiac fibrosis, cardiac myofibroblast, EZH1, EZH2, Pathology Section Received : August 31, 2016 Accepted : October 28, 2016 Published : November 03, 2016 Abstract The role of microRNA-214-3p (miR-214-3p) in cardiac fibrosis was not well illustrated. The present study aimed to investigate the expression and potential target of miR-214-3p in angiotensin II (Ang-II)-induced cardiac fibrosis. MiR-214-3p was markedly decreased in the fibrotic myocardium of a mouse Ang-II infusion model, but was upregulated in Ang-II-treated mouse myofibroblasts. Cardiac fibrosis was shown attenuated in Ang-II-infused mice received tail vein injection of miR-214-3p agomir. Consistently, miR-214-3p inhibited the expression of Col1a1 and Col3a1 in mouse myofibroblasts in vitro . MiR-214-3p could bind the 3’-UTRs of enhancer of zeste homolog 1 (EZH1) and -2, and suppressed EZH1 and -2 expressions at the transcriptional level. Functionally, miR-214-3p mimic, in parallel to EZH1 siRNA and EZH2 siRNA, could enhance peroxisome proliferator-activated receptor-γ (PPAR-γ) expression and inhibited the expression of Col1a1 and Col3a1 in myofibroblasts. In addition, enforced expression of EZH1 and -2, and knockdown of PPAR-γ resulted in the increase of Col1a1 and Col3a1 in myofibroblasts. Moreover, the NF-κB signal pathway was verified to mediate Ang-II-induced miR-214-3p expression in myofibroblasts. Taken together, our results revealed that EZH1 and -2 were novel targets of miR-214-3p, and miR-214-3p might be one potential miRNA for the prevention of cardiac fibrosis.