Vascular Endothelial Growth Factor 165 (VEGF165) Activities Are Inhibited by Carboxymethyl Benzylamide Dextran that Competes for Heparin Binding to VEGF165 and VEGF165·KDR Complexes

Yamina Hamma-Kourbali, Roger Vassy, Anna Starzec, Valérie Le Meuth-Metzinger, Olivier Oudar, Rozita Bagheri-Yarmand, Gérard Perret, Michel Crépin

Research output: Contribution to journalArticle

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Abstract

We have previously shown that carboxymethyl dextran benzylamide (CMDB7), a heparin-like molecule, inhibits the growth of tumors xenografted in nude mice, angiogenesis, and metastasis by altering the binding of angiogenic growth factors, including platelet-derived growth factor, transforming growth factor β, and fibroblast growth factor 2, to their specific receptors. In this study, we explore the effect of CMDB7 on the most specific angiogenic growth factor, vascular endothelial growth factor 165 (VEGF165). We demonstrate here that CMDB7 inhibits the mitogenic effect of VEGF165 on human umbilical vein endothelial cells (HUV-ECs) by preventing the VEGF 165-induced VEGF receptor-2 (KDR) autophosphorylation and consequently a specific intracellular signaling. In competition experiments, the binding of 125I-VEGF165 to HUV-ECs is inhibited by CMDB7 with an IC50 of 2 μM. Accordingly, CMDB7 inhibits the cross-linking of 125I-VEGF165 to the surface of HUV-ECs, causing the disappearance of both labeled complexes, 170-180 and 240-250 kDa. We show that CMDB7 increases the electrophoretic mobility of VEGF 165, thus evidencing formation of a stable complex with this factor. Moreover, CMDB7 reduces the 125I-VEGF165 binding to coated heparin-albumin and prevents a heparin-induced increase in iodinated VEGF165 binding to soluble 125I-KDR-Fc chimera. Concerning KDR, CMDB7 has no effect on 125I-KDR-Fc electrophoretic migration and does not affect labeled KDR-Fc binding to coated heparin-albumin. In the presence of VEGF165, 125I-KDR-Fc binding to heparin is enhanced, and under these conditions, CMDB7 interferes with KDR binding. These data indicate that CMDB7 effectively inhibits the VEGF 165 activities by interfering with heparin binding to VEGF 165 and VEGF165·KDR complexes but not by direct interactions with KDR.

Original languageEnglish (US)
Pages (from-to)39748-39754
Number of pages7
JournalJournal of Biological Chemistry
Volume276
Issue number43
DOIs
StatePublished - Oct 26 2001

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Dextrans
Heparin
Endothelial cells
Human Umbilical Vein Endothelial Cells
Angiogenesis Inducing Agents
Albumins
Intercellular Signaling Peptides and Proteins
human VEGFA protein
carboxymethyl dextran
CMDBS 25
Vascular Endothelial Growth Factor Receptor-2
Electrophoretic mobility
Vascular Endothelial Growth Factor Receptor
Platelet-Derived Growth Factor
Transforming Growth Factors
Fibroblast Growth Factor 2
Nude Mice
Inhibitory Concentration 50
Tumors
Neoplasm Metastasis

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Vascular Endothelial Growth Factor 165 (VEGF165) Activities Are Inhibited by Carboxymethyl Benzylamide Dextran that Competes for Heparin Binding to VEGF165 and VEGF165·KDR Complexes. / Hamma-Kourbali, Yamina; Vassy, Roger; Starzec, Anna; Le Meuth-Metzinger, Valérie; Oudar, Olivier; Bagheri-Yarmand, Rozita; Perret, Gérard; Crépin, Michel.

In: Journal of Biological Chemistry, Vol. 276, No. 43, 26.10.2001, p. 39748-39754.

Research output: Contribution to journalArticle

Hamma-Kourbali, Yamina ; Vassy, Roger ; Starzec, Anna ; Le Meuth-Metzinger, Valérie ; Oudar, Olivier ; Bagheri-Yarmand, Rozita ; Perret, Gérard ; Crépin, Michel. / Vascular Endothelial Growth Factor 165 (VEGF165) Activities Are Inhibited by Carboxymethyl Benzylamide Dextran that Competes for Heparin Binding to VEGF165 and VEGF165·KDR Complexes. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 43. pp. 39748-39754.
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abstract = "We have previously shown that carboxymethyl dextran benzylamide (CMDB7), a heparin-like molecule, inhibits the growth of tumors xenografted in nude mice, angiogenesis, and metastasis by altering the binding of angiogenic growth factors, including platelet-derived growth factor, transforming growth factor β, and fibroblast growth factor 2, to their specific receptors. In this study, we explore the effect of CMDB7 on the most specific angiogenic growth factor, vascular endothelial growth factor 165 (VEGF165). We demonstrate here that CMDB7 inhibits the mitogenic effect of VEGF165 on human umbilical vein endothelial cells (HUV-ECs) by preventing the VEGF 165-induced VEGF receptor-2 (KDR) autophosphorylation and consequently a specific intracellular signaling. In competition experiments, the binding of 125I-VEGF165 to HUV-ECs is inhibited by CMDB7 with an IC50 of 2 μM. Accordingly, CMDB7 inhibits the cross-linking of 125I-VEGF165 to the surface of HUV-ECs, causing the disappearance of both labeled complexes, 170-180 and 240-250 kDa. We show that CMDB7 increases the electrophoretic mobility of VEGF 165, thus evidencing formation of a stable complex with this factor. Moreover, CMDB7 reduces the 125I-VEGF165 binding to coated heparin-albumin and prevents a heparin-induced increase in iodinated VEGF165 binding to soluble 125I-KDR-Fc chimera. Concerning KDR, CMDB7 has no effect on 125I-KDR-Fc electrophoretic migration and does not affect labeled KDR-Fc binding to coated heparin-albumin. In the presence of VEGF165, 125I-KDR-Fc binding to heparin is enhanced, and under these conditions, CMDB7 interferes with KDR binding. These data indicate that CMDB7 effectively inhibits the VEGF 165 activities by interfering with heparin binding to VEGF 165 and VEGF165·KDR complexes but not by direct interactions with KDR.",
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AU - Vassy, Roger

AU - Starzec, Anna

AU - Le Meuth-Metzinger, Valérie

AU - Oudar, Olivier

AU - Bagheri-Yarmand, Rozita

AU - Perret, Gérard

AU - Crépin, Michel

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AB - We have previously shown that carboxymethyl dextran benzylamide (CMDB7), a heparin-like molecule, inhibits the growth of tumors xenografted in nude mice, angiogenesis, and metastasis by altering the binding of angiogenic growth factors, including platelet-derived growth factor, transforming growth factor β, and fibroblast growth factor 2, to their specific receptors. In this study, we explore the effect of CMDB7 on the most specific angiogenic growth factor, vascular endothelial growth factor 165 (VEGF165). We demonstrate here that CMDB7 inhibits the mitogenic effect of VEGF165 on human umbilical vein endothelial cells (HUV-ECs) by preventing the VEGF 165-induced VEGF receptor-2 (KDR) autophosphorylation and consequently a specific intracellular signaling. In competition experiments, the binding of 125I-VEGF165 to HUV-ECs is inhibited by CMDB7 with an IC50 of 2 μM. Accordingly, CMDB7 inhibits the cross-linking of 125I-VEGF165 to the surface of HUV-ECs, causing the disappearance of both labeled complexes, 170-180 and 240-250 kDa. We show that CMDB7 increases the electrophoretic mobility of VEGF 165, thus evidencing formation of a stable complex with this factor. Moreover, CMDB7 reduces the 125I-VEGF165 binding to coated heparin-albumin and prevents a heparin-induced increase in iodinated VEGF165 binding to soluble 125I-KDR-Fc chimera. Concerning KDR, CMDB7 has no effect on 125I-KDR-Fc electrophoretic migration and does not affect labeled KDR-Fc binding to coated heparin-albumin. In the presence of VEGF165, 125I-KDR-Fc binding to heparin is enhanced, and under these conditions, CMDB7 interferes with KDR binding. These data indicate that CMDB7 effectively inhibits the VEGF 165 activities by interfering with heparin binding to VEGF 165 and VEGF165·KDR complexes but not by direct interactions with KDR.

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