Pancreatic cancer cell and microparticle procoagulant surface characterization

Abstract

Advanced pancreatic cancer is associated with a high risk of patients developing venous thromboembolism. This increased risk is thought to be tumour-driven and associated with tissue factor (TF) and microparticles. The aim of this study was to investigate the role of TF and phospholipid expression in the procoagulant properties of pancreatic cell lines and microparticles. Pancreatic cancer cell lines (MIA-PaCa-2, ASPC-1 and CFPAC-1) were assessed for expression of TF and microparticle release. Procoagulant potential was determined by a prothrombin time assay. Cell surface expression of TF was highest in CFPAC-1, with low expression on ASPC-1 and little/no expression on MIA-PaCa-2. Clotting time (CT) was cell number and TF-dependent (P < 0.001). Blocking of TF resulted in slower CT for CFPAC-1 and ASPC1 and prevented clotting in MIA-PaCa-2. Microparticles were shown to be procoagulant and the majority of procoagulant potential could be removed by passing cell-free media through a 0.1 μm filter. A dose-dependent CT was observed in both ASPC-1 and CFPAC-1 cell-free media. Furthermore, addition of duramycin prevented microparticle-supported coagulation. The data presented suggest a key role for cell and microparticle surface-expressed TF and phospholipids in coagulation and highlight duramycin-mediated disruption of clotting.

Evaluation of coagulation abnormalities in acute liver failure

“…Microparticles derived from monocytes are also known to carry tissue factor and are able to initiate and support coagulation [15]. An increase in microparticles has been demonstrated in various thrombotic scenarios such as acute coronary syndrome and cancer [16,17]. However, here, we show for the first time that this is true in ALF as well, thus contributing to thrombotic potential in these patients.…”

Anticancer effects of monocarbonyl analogs of curcumin: oxidative stress, nuclear translocation and modulation of AP-1 and NF-κB

“…PDA pathobiology is marked by high exposure of surface PS and up-regulation of full-length Tissue Factor (flTF), an obligatory enzymatic cofactor for serine protease fVIIa and the main trigger of blood clotting which, together with high surface PS levels, is believed to be major contributing factors to high rates of thrombosis in PDA. 57 We recently showed that a minimally coagulant alternatively spliced form of TF (asTF), also expressed at high levels in PDA lesions, promotes tumor growth and spread non-proteolytically, acting as an integrin ligand. 52 Because both flTF and asTF contribute to tumor growth and spread, as does PS, we sought to examine whether EF24 and/or UBS109 exhibit cytotoxic activity against four human PDA lines: Mia-PaCa-2 (high surface PS, no fl/asTF); ASPC-1(low surface PS, more flTF than asTF); Pt45P1 (medium surface PS, more flTF than asTF), and Pt45P1/asTF+ (medium surface PS, flTF approximately equals asTF).…”

Development of Coagulation Factor Probes for the Identification of Procoagulant Circulating Tumor Cells

“…The ability of cancer cell lines to clot plasma is abrogated by incubating with a TF-blocking antibody, or with Annexin V, which blocks the binding of coagulation factors to the PS-containing cancer cell membrane (Berny-Lang et al, 2011). Further, the clotting kinetics for plasma spiked with cancer cells is strongly dependent upon the number of cells added (Tormoen et al, 2011; Yates et al, 2011; Welsh et al, 2012). Therefore, it appears that cancer cells are wholly capable of cell-mediated coagulation in vitro , whereby they can initiate coagulation through surface expression of TF and facilitate the propagation of coagulation by binding and assembling coagulation factor complexes upon their cell membranes.…”