Overexpression of miR-126 inhibits the activation and migration of HSCs through targeting CRK

Cell Physiol Biochem. 2014;33(1):97-106. doi: 10.1159/000356653. Epub 2014 Jan 17.

Abstract

Background & aims: MicroRNAs (miRNAs) have been shown to play essential roles in HSCs activation which contributes to hepatic fibrosis. Our previous miRNA microarray results suggested that miR-126 might be decreased during HSCs activation as other studies. The aim of this study is to investigate the role of miR-126 during HSCs activation.

Methods: In this study, the expression of miR-126 during HSCs activation was measured and confirmed by qRT-PCR. Then, miR-126 expression was restored by transfection of lentivirus vector encoding miR-126. Futhermore, cell proliferation was assayed by the cell counting kit-8 (CCK-8), cell migration was assayed by transwell assay, and the markers of activation of HSCs, α-SMA and collagen type I, were assayed by qRT-PCR, Western Blotting, Immunostaining and ELISA. Luciferase reporter assay was used to find the target of miR-126, and Western Blotting and Immunostaining was used to validate the target of miR-126. Then, the expression and the role of the target of miR-126 during HSCs activation was further assessed.

Results: The expression of miR-126 was confirmed to be significantly decreased during HSCs activation. Overexpression of miR-126 significantly inhibited HSCs migration but did not affect HSCs proliferation. The expression of α-SMA and collagen type I were both obviously decreased by miR-126 restoration. CRK was found to be the target of miR-126 and overexpression of miR-126 significantly inhibited CRK expression. And it was found that overexpression of CRK also significantly decreased miR-126 expression and promoted HSCs activation.

Conclusions: Our study showed that overexpression of miR-126 significantly inhibited the activation and migration of HSCs through targeting CRK which can also decrease miR-126 expression and promote HSCs activation.

MeSH terms

  • Animals
  • Base Sequence
  • Cell Line
  • Cell Movement*
  • Cell Proliferation
  • Collagen Type I / metabolism
  • Gene Expression Regulation
  • Hepatic Stellate Cells / cytology*
  • Hepatic Stellate Cells / metabolism*
  • Male
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • Molecular Sequence Data
  • Proto-Oncogene Proteins c-crk / metabolism*
  • Rats, Sprague-Dawley

Substances

  • Collagen Type I
  • Crk protein, rat
  • MIRN126 microRNA, rat
  • MicroRNAs
  • Proto-Oncogene Proteins c-crk