Mechanism for diminished tissue factor expression by endothelial cells cultured with heparin binding growth factor-1 and heparin.
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.
Almus
F E
FE
Department of Medicine, University of California, San Diego, La Jolla.
Rao
L V
LV
Pendurthi
U R
UR
Quattrochi
L
L
Rapaport
S I
SI
D000941:Antigens / Q000276:immunology; D001059:Apoproteins / Q000276:immunology; D015152:Blotting, Northern; D002452:Cell Count / Q000187:drug effects; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D004247:DNA / Q000235:genetics; D015342:DNA Probes; D004730:Endothelium, Vascular / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D016220:Fibroblast Growth Factor 1 / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects; D006493:Heparin / Q000494:pharmacology*; D006801:Humans; D010743:Phospholipids / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013917:Thrombin / Q000494:pharmacology; D013925:Thromboplastin / Q000235:genetics / Q000378:metabolism*
D000941:Antigens; D001059:Apoproteins; D015342:DNA Probes; D010743:Phospholipids; D012333:RNA, Messenger; D016220:Fibroblast Growth Factor 1; D006493:Heparin; D004247:DNA; D013925:Thromboplastin; D013917:Thrombin