MicroRNA-93 regulates collagen loss by targeting MMP3 in human nucleus pulposus cells

Cell Prolif. 2015 Jun;48(3):284-92. doi: 10.1111/cpr.12176. Epub 2015 Mar 27.

Abstract

Objectives: Degenerated disc disease is one of the most common medical conditions in patients suffering from low back pain. Recent studies have shown that microRNAs can regulate cell function in many pathological conditions. The aim of this study was to investigate expression and role of miR-93 in disc degeneration.

Materials and methods: Quantitative RT-PCR was employed to investigate level of miR-93 in degenerative nucleus pulposus (NP) tissues. Then, functional analysis of miR-93 in regulating collagen II expression was performed. Subsequently, western blotting and luciferase reporter assay were used to detect the target gene.

Results: We showed that miR-93 was significantly down-regulated in degenerative NP tissues and its levels were associated with grade of disc degeneration. Overexpression of miR-93 stimulated type II collagen expression in NP cells. Moreover, MMP3 was identified as a putative target of miR-93. MiR-93 inhibited MMP3 expression by directly targeting its 3'UTR, and this was abolished by miR-93 binding site mutations. Additionally, restoration of MMP3 in miR-93-overexpressed NP cells reversed effects of type II collagen expression. Expression of MMP3 inversely correlated with miR-93 expression in degenerative NP tissues.

Conclusions: Taken together, we demonstrated that miR-93 contributed to abnormal NP cell type II collagen expression by targeting MMP3, involved in intervertebral disc degeneration.

MeSH terms

  • 3' Untranslated Regions
  • Adolescent
  • Adult
  • Apoptosis
  • Binding Sites
  • Cell Differentiation
  • Cell Movement
  • Cell Proliferation
  • Chondrocytes / metabolism*
  • Chondrocytes / pathology
  • Collagen Type II / genetics*
  • Collagen Type II / metabolism
  • Epigenesis, Genetic
  • Female
  • Genes, Reporter
  • Humans
  • Intervertebral Disc / metabolism*
  • Intervertebral Disc / pathology
  • Intervertebral Disc Degeneration / genetics*
  • Intervertebral Disc Degeneration / metabolism
  • Intervertebral Disc Degeneration / pathology
  • Luciferases / genetics
  • Luciferases / metabolism
  • Male
  • Matrix Metalloproteinase 3 / genetics*
  • Matrix Metalloproteinase 3 / metabolism
  • MicroRNAs / genetics*
  • MicroRNAs / metabolism
  • Middle Aged
  • Mutation
  • Real-Time Polymerase Chain Reaction
  • Severity of Illness Index
  • Signal Transduction

Substances

  • 3' Untranslated Regions
  • Collagen Type II
  • MIRN93 microRNA, human
  • MicroRNAs
  • Luciferases
  • MMP3 protein, human
  • Matrix Metalloproteinase 3