Specific and non-specific KGF inhibition of KGF-induced breast cancer cell motility

Anticancer Res. Sep-Oct 2002;22(5):2539-45.

Abstract

Background: Keratinocyte growth factor (KGF), a member of the fibroblast growth factor family, is a mesenchymally derived mediator of epithelial cell proliferation and migration. In a previous study, we reported that KGF enhanced the motility of human breast cancer cells. The objective of the present study was to examine the influence of specific and non-specific KGF inhibitors on KGF-induced motility and proliferation in ER-positive MCF-7 cells.

Materials and methods: In the present study three KGF inhibitors were employed [Heparin, Innohep, a low molecular weight heparin (LMWH) and KGFR2 beta (IIIb)/Fc, a chimeric KGFR fragment]. Heparin and LMWH bind to low affinity sites on KGF and produce non-specific inhibition, while KGFR2 beta (IIIb)/Fc, a soluble chimera of an extracellular KGFR fragment, is a more specific KGF inhibitor. Cellular motility was measured using two methods: culture wounding over a period of 48 hours; and secondly, time-lapse videomicroscopy (TLVM).

Results: In these experiments KGF was found to produce a dose-dependent enhancement of MCF-7 cell motility over a dosage range of 5 to 500 ng/ml. In the TLVM experiments, Heparin (30 ng/ml), LMWH (30 ng/ml) and KGFR2 beta (IIIb)/Fc (50 micrograms/ml) completely inhibited KGF-induced motility of MCF-7 cells during the initial 2-hour observation period. In the culture wounding assay, LMWH produced a greater reduction in KGF-induced motility than heparin at 48 hours post-treatment.

Conclusion: The results of this study indicate that KGF-mediated enhancement of breast cancer cells motility and proliferation is inhibited by both specific and non-specific KGF inhibitors. LMWH appears to produce an inhibition of KGF with a much longer duration of action than Heparin. Our results suggest that KGF inhibition may be a potential new therapeutic approach for the treatment of metastatic breast cancer.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Breast Neoplasms / pathology*
  • Cell Division / drug effects
  • Cell Division / physiology
  • Cell Movement / drug effects*
  • Cell Movement / physiology
  • Dose-Response Relationship, Drug
  • Fibroblast Growth Factor 7
  • Fibroblast Growth Factors / antagonists & inhibitors*
  • Fibroblast Growth Factors / pharmacology
  • Heparin / pharmacology
  • Heparin, Low-Molecular-Weight / pharmacology
  • Immunoglobulin Fc Fragments / pharmacology
  • Microscopy, Video
  • Peptide Fragments / pharmacology
  • Receptor, Fibroblast Growth Factor, Type 2
  • Receptors, Fibroblast Growth Factor / chemistry
  • Recombinant Fusion Proteins / pharmacology
  • Recombinant Proteins / antagonists & inhibitors
  • Recombinant Proteins / pharmacology
  • Substrate Specificity
  • Tumor Cells, Cultured

Substances

  • Heparin, Low-Molecular-Weight
  • Immunoglobulin Fc Fragments
  • Peptide Fragments
  • Receptors, Fibroblast Growth Factor
  • Recombinant Fusion Proteins
  • Recombinant Proteins
  • Fibroblast Growth Factor 7
  • Fibroblast Growth Factors
  • Heparin
  • Receptor, Fibroblast Growth Factor, Type 2
  • keratinocyte growth factor receptor