MiR-9a-5p regulates proliferation and migration of hepatic stellate cells under pressure through inhibition of Sirt1

• Published in: World journal of gastroenterology : WJG 2015-09, Vol.21 (34), p.9900-9915
• Main Authors: Qi, Feng, Hu, Jiang-Feng, Liu, Bao-Hai, Wu, Chao-Qun, Yu, Hong-Yu, Yao, Ding-Kang, Zhu, Liang
• Format: Article
• Language: English
• Subjects: Animals; Base Sequence; Basic Study; Cell Movement; Cell Proliferation; Cells, Cultured; Gene Expression Profiling - methods; Genes, Reporter; Hepatic Stellate Cells - metabolism; Hepatic Stellate Cells - pathology; Hypertension, Portal - genetics; Hypertension, Portal - metabolism; Hypertension, Portal - pathology; Hypertension, Portal - physiopathology; Liver Cirrhosis, Experimental - complications; Male; Mechanotransduction, Cellular; MicroRNAs - genetics; MicroRNAs - metabolism; Molecular Sequence Data; Oligonucleotide Array Sequence Analysis; Portal Pressure; Primary Cell Culture; Rats; Rats, Sprague-Dawley; Real-Time Polymerase Chain Reaction; Sirtuin 1 - genetics; Sirtuin 1 - metabolism; Time Factors; Transfection
• ISSN: 1007-9327; 2219-2840
• DOI: 10.3748/wjg.v21.i34.9900

To reveal the functions of microRNAs (miRNAs) with respect to hepatic stellate cells (HSCs) in response to portal hypertension. Primary rat HSCs were exposed to static water pressure (10 mmHg, 1 h) and the pressure-induced miRNA expression profile was detected by next-generation sequencing. Quantitative real-time polymerase chain reaction was used to verify the expression of miRNAs. A potential target of MiR-9a-5p was measured by a luciferase reporter assay and Western blot. CCK-8 assay and Transwell assay were used to detect the proliferation and migration of HSCs under pressure. According to the profile, the expression of miR-9a-5p was further confirmed to be significantly increased after pressure overload in HSCs (3.70 ± 0.61 vs 0.97 ± 0.15, P = 0.0226), which resulted in the proliferation, migration and activation of HSCs. In vivo, the up-regulation of miR-9a-5p (2.09 ± 0.91 vs 4.27 ± 1.74, P = 0.0025) and the down-regulation of Sirt1 (2.41 ± 0.51 vs 1.13 ± 0.11, P = 0.0006) were observed in rat fibrotic liver with portal hypertension. Sirt1 was a potential target gene of miR-9a-5p. Through restoring the expression of Sirt1 in miR-9a-5p transfected HSCs on pressure overload, we found that overexpression of Sirt1 could partially abrogate the miR-9a-5p mediated suppression of the proliferation, migration and activation of HSCs. Our results suggest that during liver fibrosis, portal hypertension may induce the proliferation, migration and activation of HSCs through the up-regulation of miR-9a-5p, which targets Sirt1.