Upregulation of miR-146a-5p is associated with increased proliferation and migration of vascular smooth muscle cells in aortic dissection

J Clin Lab Anal. 2019 May;33(4):e22843. doi: 10.1002/jcla.22843. Epub 2019 Feb 19.

Abstract

Background: This study aimed to investigate whether miR-146a-5p was involved in the pathogenesis of thoracic aortic dissection (AD) via regulating the biological function of vascular smooth muscle cells (VSMCs).

Methods: Circulating miR-146a-5p level was measured by quantitative polymerase chain reaction (qPCR) in AD patients and healthy controls. Human dissected aortic samples were obtained from patients with thoracic AD Stanford type A undergoing surgical repair, and normal control samples were from organ donors who died from nonvascular diseases. The expression level of miR-146a-5p was detected using qPCR in each sample. The expression of SMAD4, which is involved in the TGF-β pathway and indicated as the target gene of miR-146a-5p, was measured by qPCR and Western blot analysis at the mRNA level and protein level, respectively. Subsequently, VSMCs were transfected with miR-146a-5p mimics or inhibitors in vitro. VSMC proliferation and migration were detected using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and Transwell assay, respectively. Flow cytometry was used to identify apoptosis. The expression of SMAD4 in VSMCs was determined using qPCR and Western blot analysis.

Results: Plasma level of miR-146a-5p is significantly higher in the AD group as compared with the control group. The expression of miR-146a-5p was significantly upregulated in dissected aorta compared with controls (P < 0.05). The overexpression of miR-146a-5p significantly induced VSMC proliferation and migration in vitro.

Conclusions: The expression of SMAD4 was modulated by miR-146a-5p. miR-146a-5p induced VSMC proliferation and migration through targeting SMAD4 and hence might be potentially involved in the development of AD.

Keywords: SMAD4; aortic dissection; miR-146a-5p; migration; proliferation; vascular smooth muscle cell.

MeSH terms

  • Adult
  • Aortic Aneurysm, Thoracic / genetics
  • Aortic Aneurysm, Thoracic / pathology*
  • Aortic Dissection / genetics
  • Aortic Dissection / pathology*
  • Case-Control Studies
  • Cell Movement / genetics
  • Cell Proliferation / genetics
  • Cell-Free Nucleic Acids / blood
  • Cells, Cultured
  • Female
  • Gene Expression Regulation
  • Humans
  • Male
  • MicroRNAs / blood
  • MicroRNAs / genetics*
  • Middle Aged
  • Muscle, Smooth, Vascular / pathology*
  • Smad4 Protein / genetics*
  • Smad4 Protein / metabolism
  • Up-Regulation

Substances

  • Cell-Free Nucleic Acids
  • MIRN146 microRNA, human
  • MicroRNAs
  • SMAD4 protein, human
  • Smad4 Protein