Assembly and regulation of prothrombinase complex on B16F10 melanoma cells.
A number of studies indicate that coagulation proteases play significant roles in cancer biology. Melanoma is a highly metastatic cancer, and there is evidence that thrombin contributes to this aggressive pattern. However, few studies correlate this type of cancer with formation of the prothrombinase complex, which is responsible for conversion of prothrombin into thrombin in the coagulation system. The aim of this study was to investigate the assembly and regulation of prothrombinase complex on the murine melanoma cell line, B16F10. B16F10 cells were unable to activate prothrombin except when previously incubated with factor Xa. This effect was dependent on factor Xa binding to cell membranes, since no activation was detected with Gla-domainless factor Xa. The thrombin formation by B16F10-bound factor Xa was enhanced approximately 10 fold in the presence of factor Va, indicating the assembly of prothrombinase complex. Differently from platelets, B16F10-assembled prothrombinase complex was inhibited by prothrombin fragment 1 but not by fragment 2. In addition, bothrojaracin, a specific ligand of proexosite I on prothrombin, caused a significant decrease in the zymogen activation. Our data demonstrate that B16F10 melanoma cells generate thrombin by promoting assembly of the prothrombinase complex. This ability might be correlated with the increased metastatic potential of this cell line. Moreover, B16F10-assembled prothrombinase complex seems to be modulated in a different way from that found for the physiological complex assembled on platelets.
Kirszberg
Clarice
C
Departamento de Bioquímica Médica, Instituto de Ciências Biomédicas, Centro de Ciências da Saúde, Universidade Federal do Rio de Janeiro, Cidade Universitária, Ilha do Fundão, Avenida Bauhínia 400, Rio de Janeiro, CEP-21941-590, Rio de Janeiro, Brazil.
Rumjanek
Vivian M
VM
Monteiro
Robson Q
RQ
D000818:Animals; D001777:Blood Coagulation; D045744:Cell Line, Tumor; D003435:Crotalid Venoms / Q000494:pharmacology; D005165:Factor V / Q000096:biosynthesis* / Q000378:metabolism; D015951:Factor Xa / Q000096:biosynthesis* / Q000378:metabolism; D006801:Humans; D008545:Melanoma / Q000201:enzymology* / Q000473:pathology; D051379:Mice; D009362:Neoplasm Metastasis; D010446:Peptide Fragments / Q000494:pharmacology; D011516:Prothrombin / Q000494:pharmacology; D013917:Thrombin / Q000096:biosynthesis
D003435:Crotalid Venoms; D010446:Peptide Fragments; C045879:prothrombin fragment 1.2; C022475:prothrombinase complex; C083739:bothrojaracin; D005165:Factor V; D011516:Prothrombin; D013917:Thrombin; D015951:Factor Xa