Fibroblast activation protein increases metastatic potential of fibrosarcoma line HT1080 through upregulation of integrin-mediated signaling pathways

Clin Exp Metastasis. 2015 Jun;32(5):507-16. doi: 10.1007/s10585-015-9723-4. Epub 2015 May 21.

Abstract

The serine protease fibroblast activation protein (FAP) is selectively expressed on tumour-associated fibroblasts in most human epithelial tumours, as well as on some mesenchymal tumours such as sarcoma. High FAP expression is most often associated with poor outcome and increased metastasis. Here, we compare the in vitro metastatic potential of HT1080 fibrosarcoma cells with and without FAP expression in order to elucidate the mechanism by which FAP may influence metastasis. In the presence of FAP, cells were more adhesive to extracellular matrix proteins and migrated and invaded through Matrigel to a greater degree. The anti-FAP antibody ESC11, which caused internalization of FAP, decreased adhesion and migration, but only when cells expressed FAP. It was also found that blocking activity of integrins β1 and αvβ3 reduced both cell adhesion and migration and this effect was much more marked in FAP-expressing HT1080 cells than mock-transfected HT1080 cells. The expression or activation of intracellular proteins that form part of the downstream signaling of integrins, including integrin-linked kinase, Rac1 and focal adhesion kinase, was also upregulated when FAP was expressed, suggesting that FAP not only upregulates metastatic-like cell behaviours through interaction with integrins, but also influences the intracellular signaling of integrins. This was confirmed using both PI3 kinase and Src kinase inhibitors, which decreased adhesion and migration in FAP-expressing cells, but did not affect mock-transfected HT1080 cells. FAP is therefore a useful target for anti-cancer therapy, as not only is its expression tumour-selective, but its downregulation has the potential to reduce incidence of metastasis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal / pharmacology
  • Blotting, Western
  • Cell Adhesion
  • Cell Movement*
  • Cell Proliferation*
  • Endopeptidases
  • Fibrosarcoma / metabolism*
  • Fibrosarcoma / secondary*
  • Flow Cytometry
  • Gelatinases / antagonists & inhibitors
  • Gelatinases / immunology
  • Gelatinases / metabolism*
  • Humans
  • Integrin alphaVbeta3 / metabolism*
  • Integrin beta1 / metabolism*
  • Membrane Proteins / antagonists & inhibitors
  • Membrane Proteins / immunology
  • Membrane Proteins / metabolism*
  • Microscopy, Fluorescence
  • Neoplasm Invasiveness
  • Serine Endopeptidases / immunology
  • Serine Endopeptidases / metabolism*
  • Tumor Cells, Cultured

Substances

  • Antibodies, Monoclonal
  • Integrin alphaVbeta3
  • Integrin beta1
  • Membrane Proteins
  • Endopeptidases
  • Serine Endopeptidases
  • fibroblast activation protein alpha
  • Gelatinases