MiR-23b controls TGF-β1 induced airway smooth muscle cell proliferation via direct targeting of Smad3

Pulm Pharmacol Ther. 2017 Feb:42:33-42. doi: 10.1016/j.pupt.2017.01.001. Epub 2017 Jan 4.

Abstract

Background: MicroRNAs are small yet versatile gene tuners that regulate a variety of cellular processes, including cell growth and proliferation. Here we report that miR-23b inhibited airway smooth muscle cells (ASMCs) proliferation through directly targeting of Smad3.

Methods: We obtained ASMCs by laser capture microdissection of normal and asthmatic mice lung tissues. Mice ASMCs were cultured and induced by TGF-β1. The implication between TGF-β1 and miR-23b in ASMCs were detected by RT-PCR. The effects of miR-23b on ASMCs proliferation and apoptosis were assessed by transient transfection of miR-23b mimics and inhibitor. The expression of Smad3 in ASMCs were detected by RT-PCR and Western blotting analysis. Dual-Luciferase Reporter Assay System will be applied to identify whether Smad3 is a target gene of miR-23b.

Results: TGF-β1 and miR-23b mRNA expression of in-situ bronchial ASMCs collected by laser capture microdissection were increased in asthmatic mice compared to non-asthma controls. This is accompanied by an increase in miR-23b mRNA expression in TGF-β1 induced ASMCs. miR-23b up-regulation significantly inhibited TGF-β1-induced ASMCs proliferation and promoted apoptosis. MiR-23b negatively regulates the expression of Smad3 in ASMCs. Dual-Luciferase Reporter Assay System demonstrated that Smad3 was a direct target of miR-23b.

Conclusions: MiR-23b may function as an inhibitor of asthma airway remodeling by suppressing ASMCs proliferation via direct targeting of Smad3.

Keywords: Airway smooth muscle cells; Asthma; MiR-23b; TGF-β1/Smad3.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Airway Remodeling / genetics
  • Animals
  • Apoptosis / genetics
  • Asthma / genetics
  • Asthma / physiopathology
  • Blotting, Western
  • Cell Proliferation / genetics*
  • Disease Models, Animal
  • Female
  • Mice
  • Mice, Inbred BALB C
  • MicroRNAs / genetics*
  • Myocytes, Smooth Muscle / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Smad3 Protein / genetics*
  • Transforming Growth Factor beta1 / metabolism*
  • Up-Regulation

Substances

  • MicroRNAs
  • Mirn23b microRNA, mouse
  • Smad3 Protein
  • Transforming Growth Factor beta1