Vascular endothelial growth factor induces tissue factor and matrix metalloproteinase production in endothelial cells: conversion of prothrombin to thrombin results in progelatinase A activation and cell proliferation

Int J Cancer. 1998 Mar 2;75(5):780-6. doi: 10.1002/(sici)1097-0215(19980302)75:5<780::aid-ijc19>3.0.co;2-a.

Abstract

Production of vascular endothelial growth factor (VEGF) by cancer cells at invasive and metastatic sites is an important aspect of tumor angiogenesis. Although known primarily as a mitogen and a vascular permeability factor (VPF) for endothelial cells, VEGF/VPF has been proposed to induce the expression of procoagulant factors in endothelial cells. In this study, we have explored the ramifications of VEGF induction of tissue factor (TF) in human umbilical vein endothelial cells (HUVECs) and subsequent activation of progelatinase A. Within 3 hr of incubation with VEGF/VPF, endothelial cells accelerate TF generation as measured using chromogenic substrate assays for coagulation factors Xa and thrombin. Incubation of VEGF/VPF-pre-treated cells with prothrombin and factors X, Va, and VIIa at 37 degrees C and subsequent generation of thrombin resulted in activation of secreted endothelial progelatinase A as demonstrated by gelatin zymography. Anti-thrombin III or antibodies to TF inhibited thrombin generation and progelatinase A activation. VEGF/VPF also directly increased HUVEC secretion of interstitial collagenase, tissue inhibitor of metalloproteinases (TIMP-1) and, to a lesser extent, gelatinase A. The effect of thrombin on endothelial proliferation in serum-free media was examined. Thrombin was a growth factor for HUVECs at a lower dose than that required for progelatinase A activation. Whereas TIMP-2 abrogated thrombin-induced progelatinase A activation, it had no significant effect on thrombin-induced endothelial cell growth. We propose that an early step in tumor angiogenesis involves VEGF-induced thrombin generation and increased MMP production with subsequent activation of endothelial progelatinase A and degradation of the underlying basement membrane.

Publication types

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

MeSH terms

  • Cell Division
  • Cells, Cultured
  • Collagenases / metabolism*
  • Endothelial Growth Factors / pharmacology*
  • Endothelium, Vascular / enzymology
  • Endothelium, Vascular / metabolism*
  • Enzyme Activation
  • Enzyme Precursors / metabolism*
  • Factor X / metabolism
  • Gelatinases / metabolism*
  • Humans
  • Lymphokines / pharmacology*
  • Matrix Metalloproteinase 1
  • Metalloendopeptidases / metabolism*
  • Muscle, Smooth, Vascular / metabolism
  • Neovascularization, Pathologic
  • Prothrombin / metabolism
  • Thrombin / metabolism
  • Thromboplastin / biosynthesis*
  • Tissue Inhibitor of Metalloproteinase-1 / metabolism
  • Vascular Endothelial Growth Factor A
  • Vascular Endothelial Growth Factors

Substances

  • Endothelial Growth Factors
  • Enzyme Precursors
  • Lymphokines
  • Tissue Inhibitor of Metalloproteinase-1
  • Vascular Endothelial Growth Factor A
  • Vascular Endothelial Growth Factors
  • Prothrombin
  • Factor X
  • Thromboplastin
  • Thrombin
  • Collagenases
  • Gelatinases
  • Metalloendopeptidases
  • progelatinase
  • Matrix Metalloproteinase 1