title: Role of von Willebrand factor in tumour cell-induced platelet aggregation: differential regulation by NO and prostacyclin.
abstract: 1. We have studied the effects of a novel agonist, solid-phase von Willebrand Factor (sVWF), on tumour cell-induced platelet aggregation (TCIPA). 2. Washed platelet suspensions were obtained from human blood and the effects of HT-1080 human fibrosarcoma cells and sVWF on platelets were studied using aggregometry, phase-contrast microscopy, and flow cytometry. 3. Incubation of platelets with sVWF (1.2 microg ml(-1)) and HT-1080 cells (5 x 10(3) ml(-1)) resulted in a two-phased reaction characterized first by the adhesion of platelets to sVWF, then by aggregation. 4. TCIPA in the presence of sVWF was inhibited by S-nitroso-glutathione (GSNO, 100 microM) and prostacyclin (PGI(2), 30 nM). 5. Platelet activation in the presence of tumour cells and sVWF resulted in the decreased surface expression of platelet glycoprotein (GP)Ib and up-regulation of GPIIb/IIIa receptors. 6. Pre-incubation of platelets with PGI(2) (30 nM) resulted in inhibition of sVWF-tumour cell-stimulated platelet surface expression of GPIIb/IIIa as measured by flow cytometry using antibodies directed against both non-activated and activated receptor. In contrast, GSNO (100 microM) did not affect sVWF-tumour cell-stimulated platelet surface expression of GPIIb/IIIa. 7. Flow cytometry performed with PAC-1 antibodies that bind only to the activated GPIIb/IIIa revealed that GSNO (100 microM) caused inhibition of activation of GPIIb/IIIa. 8. The inhibitors exerted no significant effects on TCIPA-mediated changes in GPIb. 9. Thus, sVWF potentiates the platelet-aggregatory activity of HT-1080 cells and these effects appear to be mediated via up-regulation of platelet GPIIb/IIIa. 10. Prostacyclin and NO inhibit TCIPA-sVWF-mediated platelet aggregation. The mechanisms of inhibition of this aggregation by PGI(2) differ from those of NO.
authors.affiliation: Department of Pharmacology, University of Alberta, Edmonton, Alberta, T6G 2H7, Canada.
authors.forename: P
authors.initials: P
authors.lastname: Jurasz
authors.forename: M W
authors.initials: MW
authors.lastname: Stewart
authors.forename: A
authors.initials: A
authors.lastname: Radomski
authors.forename: F
authors.initials: F
authors.lastname: Khadour
authors.forename: M
authors.initials: M
authors.lastname: Duszyk
authors.forename: M W
authors.initials: MW
authors.lastname: Radomski
mesh_terms: D001792:Blood Platelets / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D018920:Coculture Techniques; D011464:Epoprostenol / Q000494:pharmacology; D006801:Humans; D009569:Nitric Oxide / Q000502:physiology; D020030:Nitric Oxide Donors / Q000494:pharmacology; D015539:Platelet Activation / Q000187:drug effects; D010973:Platelet Adhesiveness / Q000187:drug effects; D010974:Platelet Aggregation / Q000187:drug effects*; D010975:Platelet Aggregation Inhibitors / Q000494:pharmacology; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000096:biosynthesis / Q000187:drug effects; D019038:Platelet Glycoprotein GPIb-IX Complex / Q000096:biosynthesis / Q000187:drug effects; D011485:Protein Binding; D026422:S-Nitrosoglutathione / Q000494:pharmacology; D013997:Time Factors; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000187:drug effects; D014841:von Willebrand Factor / Q000378:metabolism / Q000494:pharmacology*
chemical_list: D020030:Nitric Oxide Donors; D010975:Platelet Aggregation Inhibitors; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D019038:Platelet Glycoprotein GPIb-IX Complex; D014841:von Willebrand Factor; D009569:Nitric Oxide; D026422:S-Nitrosoglutathione; D011464:Epoprostenol