MicroRNA-195 Activates Hepatic Stellate Cells In Vitro by Targeting Smad7

Abstract

Background and Aim Aberrant activation of the TGF-β1/Smad pathway contributes to the activation of hepatic stellate cells (HSCs). MicroRNA-195 has been shown to regulate the activation of HSCs. The aim of this study was to investigate the role of miRNA-195 in HSCs activation. Methods A liver fibrotic rat model induced by diethylnitrosamine was established. Dual luciferase reporter assays were performed to verify that Smad7 was the target of miRNA-195. The expression levels of miR-195, Smad7, and α-SMA in HSC-T6 transfected, respectively, with miR-195 mimic, inhibitor, or control were measured by qRT-PCR. The protein expression of Smad7 was detected by Western blot analysis. Results Enhanced miR-195 and decreased Smad7 were observed in diethylnitrosamine-induced liver fibrotic rats (P < 0.05). Dual luciferase reporter assays showed that the miR-195 mimic significantly suppressed the luciferase activity of a reporter plasmid carrying the binding site of miR-195 on the 3′UTR of Smad7 (P < 0.05). The miR-195 mimics activated HSCs, further elevated miR-195 and α-SMA (P < 0.01), and reduced the Smad7 level (P < 0.05). The miR-195 inhibitors blocked the activation of HSCs, reduced the expression of miR-195 and α-SMA (P < 0.01), and upregulated the expression of Smad7 (P < 0.05). Conclusion Collectively, we demonstrated that miRNA-195 activated HSCs by targeting Smad7.

RETRACTED: microRNA-155 Modulates Hepatic Stellate Cell Proliferation, Apoptosis, and Cell Cycle Progression in Rats With Alcoholic Hepatitis via the MAPK Signaling Pathway Through Targeting SOCS1

“…After 48 hours of transfection, cells were lysed. The luciferase activity was detected on a Luminometer TD-20/20 detector (E5311, Promega, Madison, WI, USA) using a Dual-Luciferase ® Reporter Assay System kit (Promega, Madison, WI, USA) (Song et al, 2017).…”

Inhibitory effects of oxymatrine on hepatic stellate cells activation through TGF-β/miR-195/Smad signaling pathway

“…In terms of primary HSCs, Sekiya found that miR-195 levle decreased at 10th day compared with that at 1st day; when induced by interferon-B, miR-195 level increased and interferon-B could inhibit LX-2 proliferation by down-regulating cyclin-E1 [26], these results indicated that the effect of miR-195 on pHSCs was time-limited, especially in primary HSCs. Current studies have shown that miR-195 exerted regulatory effects by targeting Smad7 [27, 28], which was consistent with our previous study in HSCs [29]. More and more evidences demonstrate that miR-195 exert regulatory effects in the liver disease, so we presumed that oxymatrine could attenuate liver fibrosis via TGF-β1/miR-195/Smad signaling pathway.…”

“…U6 snRNA and GAPDH levels were detected and served as internal control for miRNA and mRNA, respectively. In accordance with previous reports [29], we calculated the relative expression levels (2 −ΔΔCt ) of target mRNA (α-SMA, Smad7) and miR-195.…”

MiR‐195 and miR‐497 suppress tumorigenesis in lung cancer by inhibiting SMURF2‐induced TGF‐β receptor I ubiquitination

“…To identify the molecular mechanism of miR‐195 and miR‐497 in cancer, we selected the SMURF2 gene among the components of TGF‐β signaling as putative targets of miR‐195 and miR‐497 using prediction databases. In fact, many studies have reported that TGF‐β signaling promotes miR‐195 and miR‐497 expression (Chen et al , ; Duan and Chen, ; Song et al , ), and miR‐195/497 targets other proteins associated with TGF‐β signaling. We analyzed the expression of the SMURF2 gene in tumor and paired normal lung tissues from primary NSCLC patients in the NCBI GEO database (http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE21933) and lung cancer cell lines.…”