A 4-deoxy analogue of N-acetyl-D-glucosamine inhibits heparan sulphate expression and growth factor binding in vitro.
Heparan sulphate (HS) is a long, linear polysaccharide, which has a basic backbone of -beta1-4GlcA-alpha1-4GlcNAc- units. The involvement of HS in many steps of tumourigenesis, including growth and angiogenesis, makes it an appealing target for cancer therapy. To target the biosynthesis of HS by interfering with its chain elongation, a 4-deoxy analogue of N-acetyl-D-glucosamine (4-deoxy-GlcNAc) was synthesized. Using immunocytochemistry and agarose gel electrophoresis it was shown that incubation with the 4-deoxysugar resulted in a dose dependent reduction of HS expression of MV3 melanoma cells, 1 mM resulting in an almost nullified HS expression. The parent sugar GlcNAc had no effect. 4-deoxysugar treated cells were viable and proliferated at the same rate as control cells. Other glycan structures appeared to be only mildly affected, as staining by various lectins was generally not or only modestly inhibited. At 1 mM of the 4-deoxysugar, the capacity of cells to bind the HS-dependent pro-angiogenic growth factors FGF-2 and VEGF was greatly compromised. Using an in vitro angiogenesis assay, 4-deoxysugar treated endothelial cells showed a sharp reduction of FGF-2-induced sprout formation. Combined, these data indicate that an inexpensive, easily synthesized, water-soluble monosaccharide analogue can interfere with HS expression and pro-angiogenic growth factor binding.
van Wijk
Xander M R
XM
Department of Biochemistry, Nijmegen Centre for Molecular Life Sciences, Radboud University Nijmegen Medical Centre, Geert Grooteplein 26-28, 6525 GA Nijmegen, The Netherlands.
Oosterhof
Arie
A
van den Broek
Sebastiaan A M W
SA
Griffioen
Arjan W
AW
ten Dam
Gerdy B
GB
Rutjes
Floris P J T
FP
van Delft
Floris L
FL
van Kuppevelt
Toin H
TH
D000117:Acetylglucosamine / Q000031:analogs & derivatives* / Q000494:pharmacology; D002455:Cell Division / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003847:Deoxyglucose / Q000031:analogs & derivatives* / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation / Q000187:drug effects; D004353:Drug Evaluation, Preclinical; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism / Q000502:physiology; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism*; D005944:Glucosamine / Q000031:analogs & derivatives* / Q000494:pharmacology; D006497:Heparan Sulfate / Q000378:metabolism*; D006801:Humans; D008954:Models, Biological; D018919:Neovascularization, Physiologic / Q000187:drug effects; D011485:Protein Binding / Q000187:drug effects; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*
D000117:Acetylglucosamine; D003847:Deoxyglucose; D016222:Fibroblast Growth Factor 2; D005944:Glucosamine; D006497:Heparan Sulfate; D042461:Vascular Endothelial Growth Factor A; C551889:4-deoxyacetylglucosamine