Heparin oligosaccharides: inhibitors of the biological activity of bFGF on Caco-2 cells

Br J Cancer. 1997;75(1):9-16. doi: 10.1038/bjc.1997.3.

Abstract

A number of growth factors, including members of the fibroblast growth factor (FGF) family - hepatocyte growth factor, vascular endothelial growth factor and heparin-binding epidermal growth factor - are dependent on heparan sulphate (HS) for biological activity mediated through their high-affinity signal-transducing receptors. This obligate requirement for HS prompted the search for antagonists of HS function that could be used as anti-growth factor drugs for the treatment of cancer. Basic FGF (bFGF) was the focus of this study. Caco-2, a human colon carcinoma cell line, was adapted to growth in serum-free medium so that investigation of its growth factor requirements for growth and migration could be performed in defined conditions (Jayson GC, Evans GS, Pemberton PW, Lobley RW, Allen T 1994, Cancer Res, 54, 5718-5723). This cell line multiplied and moved in a dose-dependent manner in response to bFGF. Here, we show that the mitogenic response to bFGF is dependent on the presence of heparan sulphate. A library of heparin oligosaccharides with uniform composition but variable length was generated [general formula [IdoA(2S)-GlcNS(6S)n], and oligosaccharides of defined lengths were tested for their ability to inhibit the biological activity of bFGF. While intact heparin and heparin-derived fragments of 12 monosaccharide units did not affect bFGF-induced cell division or bFGF-induced cell migration, octasaccharides and decasaccharides potently inhibited the bFGF-induced growth and migration responses. In particular, octasaccharides completely inhibited these biological activities at 10 microg ml-, a clinically achievable and tolerable concentration. This study shows that the length of an oligosaccharide determines its ability to block the biological activity of bFGF. The observation that the biological activity of cell-surface heparan sulphate can be antagonized in this way in a human carcinoma cell line suggests that oligosaccharides should be investigated further as anti-growth factor agents for the treatment of cancer. In addition, the results suggest that the clinical evaluation of low-molecular weight heparin (LMWH) as an anti-cancer agent might benefit from subfractionation of the LMWH, to remove oligosaccharides of 12 or more residues.

MeSH terms

  • Animals
  • Caco-2 Cells
  • Cattle
  • Cell Division / drug effects
  • Cell Movement / drug effects
  • Chlorates / pharmacology
  • Chromatography, Gel
  • Fibroblast Growth Factor 2 / antagonists & inhibitors*
  • Fibroblast Growth Factor 2 / pharmacology
  • Glycosaminoglycans / metabolism
  • Heparin / analogs & derivatives*
  • Heparin / chemistry
  • Heparitin Sulfate / pharmacology*
  • Humans
  • Oligosaccharides / chemical synthesis
  • Oligosaccharides / isolation & purification
  • Oligosaccharides / pharmacology*

Substances

  • Chlorates
  • Glycosaminoglycans
  • Oligosaccharides
  • Fibroblast Growth Factor 2
  • Heparin
  • Heparitin Sulfate