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Mechanism for diminished tissue factor expression by endothelial cells
cultured with heparin binding growth factor-1 and heparin
FE Almus, LV Rao, UR Pendurthi, L Quattrochi and SI Rapaport
Department of Medicine, University of California, San Diego, La Jolla.
We have extended our earlier observation that growing primary cultures of
human umbilical vein endothelial cells (HUVEC) with heparin binding growth
factor 1 (HBGF-1) 20 micrograms/mL and heparin 12 U/mL inhibits expression
of tissue factor (TF) activity on HUVC monolayers perturbed with thrombin.
TF activity was measured as the ability of monolayers or cell lysates to
support FVIIa-catalyzed activation peptide release from 3H-FX. TF antigen
in HUVEC extracts was measured in an enzyme-linked immunosorbent assay
(ELISA) that uses a double-antibody sandwich technique with rabbit and goat
antibodies to human TF. TF-mRNA was measured by Northern blot hybridization
with a 32P-TF cDNA probe. Cells growth with HBGF-1/heparin had both
decreased surface and total TF activity as compared with HUVEC from the
same endothelial cell pool grown without HBGF-1/heparin. Means +/- SD for
TF antigen for four primary cultures were 4.4 +/- 0.9 ng/10(6) cells
without HBGF-1/heparin and 0.6 +/- 0.3 ng/10(6) cells with HBGF-1/heparin.
TF mRNA 4 hours after incubation with thrombin of HUVEC grown without
HBGF-1/heparin was about sevenfold higher than TF mRNA of HUVEC grown with
HBGF- 1/heparin. These data establish that growing primary cultures of
HUVEC with HBGF-1/heparin impairs their ability to synthesize TF apoprotein
after perturbation. This may be part of a generalized response of
endothelial cells to HBGF-1/heparin facilitating migration during
angiogenesis.
Volume 77,
Issue 6,
pp. 1256-1262,
03/15/1991
Copyright © 1991 by The American Society of Hematology
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