Xiaoxue Yang1, Xuelian Dan2, Ruoting Men1, Liping Ma3, Maoyao Wen1, Yong Peng4, Li Yang5. 1. Department of Gastroenterology and Hepatology, West China Hospital, West China School of Clinical Medicine, Sichuan University, Chengdu, China. 2. Department of Biomedical Sciences, City University of Hong Kong, Hong Kong, China. 3. Department of Thoracic Surgery, West China Hospital, Sichuan University, Chengdu, China. 4. Department of Thoracic Surgery, West China Hospital, Sichuan University, Chengdu, China. Electronic address: yongpeng@scu.edu.cn. 5. Department of Gastroenterology and Hepatology, West China Hospital, West China School of Clinical Medicine, Sichuan University, Chengdu, China. Electronic address: yangli_hx@scu.edu.cn.
Abstract
AIM: To understand the contribution of miR-142-3p in the activation of hepatic stellate cells (HSCs) and liver fibrosis, and the underlying mechanism. MATERIALS AND METHODS: We detected microRNAs expression profiles in quiescent and activated HSCs by microRNA-array, and performed qRT-PCR to validate these data in HSCs and plasma of cirrhosis patients. In vitro, the 3rd-5th passage HSCs was transfected with mir-142-3p mimics or stimulated with TGF β. The markers of HSCs activation (i.e. FN and α-SMA) were examined by qRT-PCR and western blotting, and cell viability was detected by MTT, colony formation assays respectively. KEY FINDING: In our study, we identified miR-142-3p as a novel regulator of HSCs activation and indicator of hepatic cirrhosis. We found that miR-142-3p was significantly reduced in activated HSCs, while TGFβRI was distinctly up-regulated in activated HSCs. Ectopic expression of miR-142-3p in activated HSCs inhibited cell viability as well as cell growth, and blocked HSCs activation, concomitant with decreased transdifferentiation markers (i.e. FN and α-SMA). Further, we confirmed that miR-142-3p was reduced upon TGF-β exposure, while diminishing TGF-β-Smad signaling pathway in turn by reducing TGFβRI expression in HSCs. Besides, the plasma level of miR-142-3p declined significantly in patients with hepatic cirrhosis. SIGNIFICANCE: In conclusion, we demonstrated that miR-142-3p repressed TGF-β-Smad signaling pathway to prevent HSCs activation through directly targeting TGFβRI in HSCs.
AIM: To understand the contribution of miR-142-3p in the activation of hepatic stellate cells (HSCs) and liver fibrosis, and the underlying mechanism. MATERIALS AND METHODS: We detected microRNAs expression profiles in quiescent and activated HSCs by microRNA-array, and performed qRT-PCR to validate these data in HSCs and plasma of cirrhosis patients. In vitro, the 3rd-5th passage HSCs was transfected with mir-142-3p mimics or stimulated with TGF β. The markers of HSCs activation (i.e. FN and α-SMA) were examined by qRT-PCR and western blotting, and cell viability was detected by MTT, colony formation assays respectively. KEY FINDING: In our study, we identified miR-142-3p as a novel regulator of HSCs activation and indicator of hepatic cirrhosis. We found that miR-142-3p was significantly reduced in activated HSCs, while TGFβRI was distinctly up-regulated in activated HSCs. Ectopic expression of miR-142-3p in activated HSCs inhibited cell viability as well as cell growth, and blocked HSCs activation, concomitant with decreased transdifferentiation markers (i.e. FN and α-SMA). Further, we confirmed that miR-142-3p was reduced upon TGF-β exposure, while diminishing TGF-β-Smad signaling pathway in turn by reducing TGFβRI expression in HSCs. Besides, the plasma level of miR-142-3p declined significantly in patients with hepatic cirrhosis. SIGNIFICANCE: In conclusion, we demonstrated that miR-142-3p repressed TGF-β-Smad signaling pathway to prevent HSCs activation through directly targeting TGFβRI in HSCs.