Down-regulation of USP13 mediates phenotype transformation of fibroblasts in idiopathic pulmonary fibrosis

Respir Res. 2015 Oct 9:16:124. doi: 10.1186/s12931-015-0286-3.

Abstract

Background: Idiopathic pulmonary fibrosis (IPF) is a fatal disease characterized by fibroblastic foci and progressive scarring of the pulmonary parenchyma. IPF fibroblasts display increased proliferation and enhanced migration and invasion, analogous to cancer cells. This transformation-like phenotype of fibroblasts plays an important role in the development of pulmonary fibrosis, but the mechanism for this is not well understood.

Methods: In this study, we compared gene expression profiles in fibrotic lung tissues from IPF patients and normal lung tissues from patients with primary spontaneous pneumothorax using a cDNA microarray to examine the mechanisms involved in the pathogenesis of IPF. In a cDNA microarray, we found that USP13 was decreased in lung tissues from patients with IPF, which was further confirmed by results from immunohistochemistry and western blot assays. Then, we used RNA interference in MRC-5 cells to inhibit USP13 and evaluated its effects by western blot, real-time RT-PCR, bromodeoxyuridine incorporation, and transwell assays. We also used co-immunoprecipitation and immunofluorescence staining to identify the correlation between USP13 and PTEN in IPF.

Results: USP13 expression levels were markedly reduced in fibroblastic foci and primary IPF fibroblast lines. The depletion of USP13 resulted in the transformation of fibroblasts into an aggressive phenotype with enhanced proliferative, migratory, and invasive capacities. Additionally, USP13 interacted with PTEN and mediated PTEN ubiquitination and degradation in lung fibroblasts.

Conclusions: Down-regulation of USP13 mediates PTEN protein loss and fibroblast phenotypic change, and thereby plays a crucial role in IPF pathogenesis.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Blotting, Western
  • Case-Control Studies
  • Cell Differentiation*
  • Cell Line
  • Cell Movement
  • Cell Proliferation
  • Down-Regulation
  • Endopeptidases / genetics
  • Endopeptidases / metabolism*
  • Epithelial-Mesenchymal Transition*
  • Fibroblasts / enzymology*
  • Fibroblasts / pathology
  • Fluorescent Antibody Technique
  • Gene Expression Profiling / methods
  • Humans
  • Idiopathic Pulmonary Fibrosis / enzymology*
  • Idiopathic Pulmonary Fibrosis / genetics
  • Idiopathic Pulmonary Fibrosis / pathology
  • Immunoprecipitation
  • Lung / enzymology*
  • Lung / pathology
  • Male
  • Middle Aged
  • Oligonucleotide Array Sequence Analysis
  • PTEN Phosphohydrolase / metabolism
  • Phenotype
  • Proteolysis
  • RNA Interference
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcriptase Polymerase Chain Reaction
  • Signal Transduction
  • Time Factors
  • Transfection
  • Ubiquitin-Specific Proteases
  • Ubiquitination

Substances

  • PTEN Phosphohydrolase
  • PTEN protein, human
  • Endopeptidases
  • USP13 protein, human
  • Ubiquitin-Specific Proteases