miR-10b-5p Regulates C2C12 Myoblasts Proliferation and Differentiation

Biosci Biotechnol Biochem. 2019 Feb;83(2):291-299. doi: 10.1080/09168451.2018.1533805. Epub 2018 Oct 18.

Abstract

The development of skeletal muscle is a complex process including myoblasts proliferation and differentiation. MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression at post-transcriptional level. Increasing evidences indicate that miRNAs are important regulators in myogenic processes. Here, we reported that the expression of miR-10b-5p steadily decreased during myoblasts proliferation, but significantly increased during myoblasts differentiation. The over-expression of miR-10b-5p promoted myoblasts proliferation and blunted myofiber formation in C2C12 cells, while miR-10b-5p down-regulation showed an opposite result. At the same time, we observed that the down-regulation of nuclear factor of activated T-cells 5 (NFAT5) repressed the differentiation of C2C12 cells, and interestingly, miR-10b-5p could suppress NFAT5 expression. Luciferase activity assays confirmed that miR-10b-5p directly target the 3'-untranslated region (3'-UTR) of NFAT5. Overall, we proposed here a novel insight that miR-10b-5p regulates the proliferation and differentiation of C2C12 myoblasts, and the impact on myogenic differentiation is partly through targeting NFAT5. Abbreviations: NFAT5: nuclear factor of activated T-cells 5; Cyclin B: cycle protein B; Cyclin D1: cycle protein D1; Cyclin E: cycle protein E; CDK4: cyclin-dependent kinase 4; MyoD: myogenic differentiation antigen; MyoG: myogenin; Myf5: myogenic factor 5; MRF4: myogenic regulatory factor 4; MyHC: myosin heavy chain; AQP5: aquaporin-5; CACNA1C: calcium voltage-gated channel subunit alpha1 C; SRF: serum response factor; Pax7: paired box 7; KLF4: Kruppel-like factor 4; 3'-UTR: 3'-untranslated region; GM: growth medium; DM: differentiation medium.

Keywords: C2C12 myoblasts; Mir-10b-5p; NFAT5; myogenesis.

MeSH terms

  • 3' Untranslated Regions
  • Animals
  • Cell Differentiation / genetics*
  • Cell Line
  • Cell Proliferation / genetics*
  • Down-Regulation
  • Humans
  • Kruppel-Like Factor 4
  • Mice
  • MicroRNAs / physiology*
  • Myoblasts / cytology*
  • Transcription Factors / genetics

Substances

  • 3' Untranslated Regions
  • KLF4 protein, human
  • Klf4 protein, mouse
  • Kruppel-Like Factor 4
  • MicroRNAs
  • NFAT5 protein, human
  • Nfat5 protein, mouse
  • Transcription Factors