INTERACTION OF VASCULOTROPIN VASCULAR ENDOTHELIAL-CELL GROWTH-FACTOR WITH HUMAN UMBILICAL VEIN ENDOTHELIAL-CELLS - BINDING, INTERNALIZATION, DEGRADATION, AND BIOLOGICAL EFFECTS

被引:121
作者
BIKFALVI, A
SAUZEAU, C
MOUKADIRI, H
MACLOUF, J
BUSSO, N
BRYCKAERT, M
PLOUET, J
TOBELEM, G
机构
[1] CTR CORDELIERS,INSERM,U86,F-75006 PARIS,FRANCE
[2] LABS GLAXO,F-91940 LES ULIS,FRANCE
[3] HOP LARIBOISIERE,INSERM,U150,F-75475 PARIS 10,FRANCE
关键词
D O I
10.1002/jcp.1041490108
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Vasculotropin/vascular endothelial cell growth factor (VAS/VEGF) is a newly purified growth factor with a unique specificity for vascular endothelial cells. We have investigated the interactions of VAS/VEGF with human umbilical vein endothelial cells (HUVE cells). I-125-VAS/VEGF was bound to HUVE cells in a saturable manner with a half-maximum binding at 2.8 ng/ml. Scatchard analysis did show two classes of high-affinity binding sites. The first class displayed a dissociation constant of 9 pM with 500 sites/cell. The dissociation constant and the number of binding sites of the second binding class were variable for different HUVE cell cultures (K(D) = 179 +/- 101 pM, 5,850 +/- 2,950 sites/cell). Half-maximal inhibition of I-125-VAS/VEGF occurred with a threefold excess of unlabeled ligand. Basic fibroblast growth factor (bFGF) and heparin did not complete with I-125-VAS/VEGF binding. In contrast, suramin and protamin sulfate completely displaced I-125-VAS/VEGF binding from HUVE cells. VAS/VEGF was shown to be internalized in HUVE cells. Maximum internalization (55% of total cell-associated radioactivity) was observed after 30 min. I-125-VAS/VEGF was completely degraded 2-3 hr after binding. At 3 hr, the trichloroacetic acid (TCA)-soluble radioactivity accumulated in the medium was 60% of the total radioactivity released by HUVE cells. No degradation fragment of I-125-VAS/VEGF was observed. Chloroquine completely inhibited degradation. VAS/VEGF was able to induce angiogenesis in vitro in HUVE cells. However, it did not significantly modulate urokinase-type plasminogen activator (u-PA), tissue-type plasminogen activator (t-PA), plasminogen activator inhibitor (PAl-1), and tissue factor (TF). Prostacyclin production was only stimulated at very high VAS/VEGF concentrations. Taken together, these results indicate that VAS/VEGF might be a potent inducer of neovascularization resulting from a direct interaction with endothelial cells. The angiogenic activity seems to be independent of the plasminogen activator or inhibitor system.
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页码:50 / 59
页数:10
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