TNFα modulates Fibroblast Growth Factor Receptor 2 gene expression through the pRB/E2F1 pathway: identification of a non-canonical E2F binding motif

PLoS One. 2013 Apr 16;8(4):e61491. doi: 10.1371/journal.pone.0061491. Print 2013.

Abstract

Interactions between epithelium and mesenchyme during wound healing are not fully understood, but Fibroblast Growth Factors (FGFs) and their receptors FGFRs are recognized as key elements. FGFR2 gene encodes for two splicing transcript variants, FGFR2-IIIb or Keratinocyte Growth Factor Receptor (KGFR) and FGFR2-IIIc, which differ for tissue localization and ligand specificity. Proinflammatory cytokines play an essential role in the regulation of epithelial-mesenchymal interactions, and have been indicated to stimulate FGFs production. Here we demonstrated that upregulation of FGFR2 mRNA and protein expression is induced by the proinflammatory cytokines Tumor Necrosis Factor-α, Interleukin-1β and Interleukin 2. Furthermore, we found that TNFα determines FGFR2 transcriptional induction through activation of pRb, mediated by Raf and/or p38 pathways, and subsequent release of the transcription factor E2F1. Experiments based on FGFR2 promoter serial deletions and site-directed mutagenesis allowed us to identify a minimal responsive element that retains the capacity to be activated by E2F1. Computational analysis indicated that this element is a non-canonical E2F responsive motif. Thus far, the molecular mechanisms of FGFR2 upregulation during wound healing or in pathological events are not known. Our data suggest that FGFR2 expression can be modulated by local recruitment of inflammatory cytokines. Furthermore, since alterations in FGFR2 expression have been linked to the pathogenesis of certain human cancers, these findings could also provide elements for diagnosis and potential targets for novel therapeutic approaches.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Western
  • Cell Line
  • Cell Line, Tumor
  • E2F1 Transcription Factor / genetics
  • E2F1 Transcription Factor / metabolism*
  • Humans
  • Immunoprecipitation
  • Microscopy, Fluorescence
  • Promoter Regions, Genetic / genetics
  • Protein Binding / drug effects
  • Real-Time Polymerase Chain Reaction
  • Receptor, Fibroblast Growth Factor, Type 2 / genetics
  • Receptor, Fibroblast Growth Factor, Type 2 / metabolism*
  • Signal Transduction / drug effects
  • Tumor Necrosis Factor-alpha / pharmacology*

Substances

  • E2F1 Transcription Factor
  • E2F1 protein, human
  • Tumor Necrosis Factor-alpha
  • Receptor, Fibroblast Growth Factor, Type 2

Grants and funding

This study was supported by grants from Regione Molise - Conv. 17/07 (B81J09001360002), Italy. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.