MiR-30c-5p regulates adventitial progenitor cells differentiation to vascular smooth muscle cells through targeting OPG

Abstract

Background As the most important component of the vascular wall, vascular smooth muscle cells (VSMCs) participate in the pathological process by phenotype transformation or differentiation from stem/progenitor cells. The main purpose of this study was to reveal the role and related molecular mechanism of microRNA-30c-5p (miR-30c-5p) in VSMC differentiation from adventitial progenitor cells expressing stem cell antigen-1(Sca-1). Methods In this study, we detected the expression of miR-30c-5p in human normal peripheral arteries and atherosclerotic arteries. In vitro, a stable differentiation model from adventitial Sca-1+ progenitor cells to VSMCs was established using transforming growth factor-β1 (TGF-β1) induction and the expression of miR-30c-5p during the process was observed. Then, we explored the effect of miR-30c-5p overexpression and inhibition on the differentiation from adventitial Sca-1+ progenitor cells to VSMCs. The target genes of miR-30c-5p were identified by protein chip and biological analyses and the expression of these genes in the differentiation process were detected. Further, the relationship between the target gene and miR-30c-5p and its effect on differentiation were evaluated. Finally, the co-transfection of miR-30c-5p inhibitor and small interfering RNA (siRNA) of the target gene was implemented to verify the functional target gene of miR-30c-5p during the differentiation from adventitial Sca-1+ progenitor cells to VSMCs, and the dual-luciferase reporter gene assay was performed to detect whether the mRNA 3′untranslated region (UTR) of the target gene is the direct binding site of miR-30c-5p. Results The expression of miR-30c-5p in the human atherosclerotic arteries was significantly lower than that in the normal arteries. During the differentiation from adventitial Sca-1+ progenitor cells to VSMCs, the expression of VSMC special markers including smooth muscle α-actin (SMαA), smooth muscle-22α (SM22α), smooth muscle myosin heavy chain (SMMHC), and h1-caponin increased accompanied with cell morphology changing from elliptic to fusiform. Meanwhile, the expression of miR-30c-5p decreased significantly. In functional experiments, overexpression of miR-30c-5p inhibited SMαA, SM22α, SMMHC, and h1-caponin at the mRNA and protein levels. In contrast, inhibition of miR-30c-5p promoted the expression of SMαA, SM22α, SMMHC, and h1-caponin. The target gene, osteoprotegerin (OPG), was predicted through protein chip and bioinformatics analyses. Overexpression of miR-30c-5p inhibited OPG expression while inhibition of miR-30c-5p had an opposite effect. Co-transfection experiments showed that low expression of OPG could weaken the promotion effect of miR-30c-5p inhibitor on the differentiation from adventitial Sca-1+ progenitor cells to VSMCs and the dual-luciferase reporter gene assay demonstrated that miR-30c-5p could target the mRNA 3′UTR of OPG directly. Conclusions This study demonstrates that miR-30c-5p expression was significantly decreased in atherosclerotic arteries and miR-30c-5p inhibited VSMC differentiation from adventitial Sca-1+ progenitor cells through targeting OPG, which may provide a new target for the treatment of VSMCs-associated diseases.

Osteoprotegerin (OPG) Upregulation Activates Breast Stromal Fibroblasts and Enhances Their Pro-Carcinogenic Effects through the STAT3/IL-6 Signaling

“…Indeed, OPG upregulation enhanced the expression of the major biomarker of active fibroblasts, a-SMA. Consistent with this, it has been previously shown that OPG positively regulates a-SMA in VSMC [ 28 ]. Likewise, OPG upregulation enhanced the expression of other active fibroblast biomarkers, namely, FAP-a, CD10, GPR77, and CD44.…”

MicroRNA-146a Promotes Embryonic Stem Cell Differentiation towards Vascular Smooth Muscle Cells through Regulation of Kruppel-like Factor 4

“…RT-qPCR was performed as described previously [27] . Briefly, RNA was extracted from the cells using Trizol (12183555, Invitrogen, USA) according to the manufacturer's instructions.…”

“…Luciferase reporter assays were performed as described in our previous studies [27] . A total of 6×10 4 ESCs were seeded in DM in a collagen-coated 12-well plate.…”

Emerging Role of MicroRNA-30c in Neurological Disorders

“…Multiple brain conditions, including ischemic stroke and thrombotic vascular events, have been linked to the overexpression of plasminogen activator inhibitor-1 (PAI-1) in the plasma, endothelial cells, and platelets [78]. MiR-30c directly targets the PAI-1 via the SERPINE1 gene [79][80][81] including the osteoprotegerin (OPG) gene [82], which mitigates atherosclerosis, thrombosis, and embolism, which are major causes of clogging of arteries [55]. MiR-30c-5p can protect against endothelial damage by targeting TGF-β1 [83] and TCF21 [58] and attenuating endothelial pyroptosis by downregulating Forkhead Box O3-NOD-, LRR-, and pyrin domain-containing protein 3 (FOXO3-NLRP3) inflammasomes [59] and α2-antiplasmin [84].…”