MicroRNA-132-3p represses Smad5 in MC3T3-E1 osteoblastic cells under cyclic tensile stress

Mol Cell Biochem. 2019 Aug;458(1-2):143-157. doi: 10.1007/s11010-019-03538-3. Epub 2019 Apr 19.

Abstract

MicroRNAs (miRNAs) regulate osteogenic differentiation of bone cells, which has applications in orthodontics. Here we evaluated the miRNA expression profile of MC3T3-E1 osteoblasts under cyclic tensile stress with chip technology and found that miR-132-3p was up-regulated by 12% cyclic tensile stress. Alkaline phosphatase activity and osteocalcin expression in MC3T3-E1 cells were decreased under these conditions. Smad2 and Smad5 were identified as potential target genes of miR-132-3p. Native and phosphorylated Smad2 and Smad5 expression was negatively correlated with miR-132-3p levels in the cells under cyclic stretch; however, only Smad5 protein level was reduced upon miR-132-3p overexpression. The luciferase reporter assay confirmed a direct interaction between miR-132-3p and Smad5. Thus, miR-132-3p maybe regulates osteoblast differentiation via Smad5 in response to cyclic tensile stress.

Keywords: Cyclic stretch; MicroRNA-132-3p; MicroRNAs; Smad5.

MeSH terms

  • Alkaline Phosphatase / genetics
  • Alkaline Phosphatase / metabolism
  • Animals
  • Cell Differentiation*
  • Cell Line
  • Mice
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • Osteoblasts / cytology
  • Osteoblasts / metabolism*
  • Smad2 Protein / genetics
  • Smad2 Protein / metabolism
  • Smad5 Protein / genetics
  • Smad5 Protein / metabolism*
  • Stress, Mechanical*
  • Tensile Strength*

Substances

  • MIRN132 microRNA, mouse
  • MicroRNAs
  • Smad2 Protein
  • Smad2 protein, mouse
  • Smad5 Protein
  • Smad5 protein, mouse
  • Alkaline Phosphatase