Involvement of miR-145 in the development of aortic dissection via inducing proliferation, migration, and apoptosis of vascular smooth muscle cells

J Clin Lab Anal. 2020 Jan;34(1):e23028. doi: 10.1002/jcla.23028. Epub 2019 Sep 6.

Abstract

Aim: The current study aimed to examine miR-145's contribution to thoracic aortic dissection (AD) development by modulating the biological functions of vascular smooth muscle cells (VSMCs).

Methods: The concentration of circulating miR-145 was determined in patients with AD and healthy controls using quantitative polymerase chain reaction (qPCR). Aortic specimens were obtained from both individuals with Stanford type A AD undergoing surgical treatment and deceased organ donors (serving as controls) whose causes of death were nonvascular diseases. Then, qPCR and fluorescence in situ hybridization were applied to assess miR-145 amounts and location, respectively. Furthermore, qPCR and immunoblot were employed to determine SMAD3 (the target gene of miR-145, involved in the TGF-β pathway) amounts at the gene and protein levels, respectively. Moreover, in vitro transfection of VSMCs with miR-145 mimics or inhibitors was conducted. Finally, the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, Transwell assay and flow cytometry were employed for detecting VSMC proliferation, migration, and apoptosis, respectively.

Results: The amounts of miR-145 in plasma and aortic specimens were markedly reduced in the AD group in comparison with control values (P < .05). miR-145 was mostly located in VSMCs. Proliferation and apoptosis of VSMCs were significantly induced in vitro by the downregulation of miR-145. Also, miR-145 modulated SMAD3 expression.

Conclusions: miR-145 was found to be downregulated in patients with AD, which induced the proliferation, migration, and apoptosis of VSMCs by targeting SMAD3. This suggested the involvement of miR-145 in the pathogenesis of AD.

Keywords: SMAD3; aortic dissection; miR-145; proliferation; vascular smooth muscle cell.

MeSH terms

  • Aged
  • Aortic Dissection / genetics*
  • Apoptosis / genetics*
  • Base Sequence
  • Cell Movement / genetics*
  • Cell Proliferation / genetics
  • Female
  • Gene Expression Regulation
  • Humans
  • Male
  • MicroRNAs / blood
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • Middle Aged
  • Muscle, Smooth, Vascular / pathology*
  • Myocytes, Smooth Muscle / metabolism*
  • Myocytes, Smooth Muscle / pathology*
  • Smad3 Protein / metabolism

Substances

  • MIRN145 microRNA, human
  • MicroRNAs
  • SMAD3 protein, human
  • Smad3 Protein