The effect of cell surface glycosaminoglycans (GAGs) on the inactivation of factor VIIa--tissue factor activity by antithrombin III.
We investigated the effect of cell surface glycosaminoglycans (GAGs) on the inactivation of factor VIIa-tissue factor activity by antithrombin III (ATIII) on a human bladder carcinoma (J82) cell line and an ovarian carcinoma (OC-2008) cell line, two tumor cell lines which constitutively synthesize and express high levels of cell surface tissue factor. We observed that ATIII inactivated factor VIIa-tissue factor more readily on OC-2008 cells than on J82 cells in the absence of added heparin. Likewise, factor Xa was more effectively inactivated on OC-2008 cells than on J82 cells. The ability of ATIII to inactivate factor VIIa-tissue factor activity on the OC-2008 cell was reduced following treatment of the cells with heparinase. This indicated that heparin-like GAGs were expressed on the OC-2008 cell surface, and that these GAGs were important for the inhibition of factor VIIa-tissue factor activity by ATIII. In addition, we demonstrated that the ability of ATIII to inactivate factor VIIa-tissue factor activity was markedly reduced following treatment of cells with calcium ionophore (A23187). However, the effect of cell surface GAGs on the inhibition of factor Xa by ATIII remained even after treatment of OC-2008 cells with A23187. In contrast to the manner of inhibition by ATIII/heparin, TFPI effectively inactivated factor VIIa-tissue factor activity on the cell surfaces even after induced physical damage or disruption of the cell by treatment with A23187. Our collective findings suggest that GAGs on cell surfaces play an important role in regulating factor VIIa-tissue factor activity by ATIII under normal conditions, or in the early phases of physical damage or destruction of the cell. However, TFPI may play a more important role than ATIII in regulating the activity of factor VIIa-tissue factor in a vascular trauma site following extensive cell injury.
Hamamoto
T
T
Department of Pathology, University of New Mexico School of Medicine, Albuquerque 87131, USA.
Kisiel
W
W
D000990:Antithrombin III / Q000378:metabolism* / Q000494:pharmacology; D001777:Blood Coagulation; D002462:Cell Membrane / Q000378:metabolism*; D015942:Factor VIIa / Q000378:metabolism*; D006025:Glycosaminoglycans / Q000378:metabolism*; D006801:Humans; D015842:Serine Proteinase Inhibitors / Q000378:metabolism* / Q000494:pharmacology; D013925:Thromboplastin / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D006025:Glycosaminoglycans; D015842:Serine Proteinase Inhibitors; D000990:Antithrombin III; D013925:Thromboplastin; D015942:Factor VIIa