miRNAs through β-ARR2/p-ERK1/2 pathway regulate the VSMC proliferation and migration

Life Sci. 2021 Aug 15:279:119703. doi: 10.1016/j.lfs.2021.119703. Epub 2021 Jun 7.

Abstract

Background: miRNAs are involved in plaque formation of atherosclerosis and vessel restenosis. In this study, we investigated the effects of miR-599, miR-204, and miR-181b on VSMC proliferation, and migration through TGFβ receptor 2 (TGFβR2), β-arrestin 2 (β-ARR2), SMAD2/p-SMAD2, and ERK1/2/p-ERK1/2.

Materials & methods: Genes and miRNAs were predicted by bioinformatics tools and were transfected by PEI-miRNAs (miR-599, miR-204, and miR-181b) complexes into VSMCs. The gene and protein expression levels were evaluated by real-time RT-PCR and western blotting techniques, respectively. The VSMC proliferation and migration were studied by MTT and scratch assay, respectively.

Results: The miR-181b and miR-204 downregulated significantly β-ARR2 gene and protein expression levels and p-ERK1/2 values. Moreover, TGFβR2 gene and protein expression levels and p-SMAD2 values were not significantly affected by miR-181b and miR-204. The VSMC proliferation (p = 0.0019, p = 0.0054, respectively) and migration (p < 0.0001, p < 0.0001, respectively) were inhibited by the miR-181b and miR-204. The miR-599 inhibited VSMC proliferation (p = 0.044) and migration (p = 0.0055) but it did not affect significantly the β-ARR2 and TGFβR2 gene and protein expression levels.

Conclusion: The results suggested that the inhibitory effects of miR-181b and miR-204 on VSMC proliferation and migration are mediated by the β-ARR2/p-ERK1/2 pathway. Since VSMC proliferation and migration are involved in plaque growth, therefore this pathway can be a therapeutic target for atherosclerosis.

Keywords: ERK1/2; Migration; SMAD2; TGFβR2; VSMCs; β-ARR2.

MeSH terms

  • Cell Movement*
  • Cell Proliferation*
  • Gene Expression Regulation*
  • Gene Regulatory Networks
  • Humans
  • MicroRNAs / genetics*
  • Mitogen-Activated Protein Kinase 1 / genetics
  • Mitogen-Activated Protein Kinase 1 / metabolism
  • Mitogen-Activated Protein Kinase 3 / genetics
  • Mitogen-Activated Protein Kinase 3 / metabolism
  • Muscle, Smooth, Vascular / cytology*
  • Muscle, Smooth, Vascular / physiology
  • Myocytes, Smooth Muscle / cytology*
  • Myocytes, Smooth Muscle / physiology
  • Signal Transduction
  • beta-Arrestin 2 / genetics
  • beta-Arrestin 2 / metabolism

Substances

  • MIRN204 microRNA, human
  • MIrn181 microRNA, human
  • MicroRNAs
  • beta-Arrestin 2
  • MAPK1 protein, human
  • MAPK3 protein, human
  • Mitogen-Activated Protein Kinase 1
  • Mitogen-Activated Protein Kinase 3