Modelization the impact of antithrombotic agents on pancreatic tumoral micro-environment.

Objective Interactions between cancer cells and their micro-environment with antithrombotic agents is an emerging field of research. In the present study we investigated the impact of apixaban, fondaparinux, enoxaparin and tinzaparin on the procoagulant properties of pancreatic cancer cells BXPC3. Reciprocally, we investigated the impact of BXPC3 on the potency of these antithrombotic agents.

Material and Methods BXPC3 (400 cells/ml) were exposed to apixaban (2 microgram/ml), fondaparinux (2 microgram/ml), enoxaparin, or tinzaparin (2 anti-Xa IU/ml) for 48h. Then, cells and supernatants were separated and added in normal platelet poor plasma (PPP) for thrombin generation (TG) experiments. Viability of cancer cells (assessed with the MTT assay), gene expression for TF, VEGF, THSB1 (assessed with RT-qPCR) and expression of TF protein and activity were also examined. Microparticles (MP) were tested for TF with specific ELISA. Residual anti-Xa activity was measured in the supernatant using specific amidolytic assays.

Results BXPC3 enhanced TG. Incubation of the BXPC3 cells with all antithrombotic agents did not significantly modify their TG capacity. Apixaban resulted in significant TF mRNA expression decrease by BXPC3 cells. None of the antithrombotic agents significantly modified the amount of BXPC3 cells -TF protein. Fondaparinux and enoxaparin significantly decreased VEGF mRNA expression and apixaban significantly increased the expression of THBS1. The viability of BXPC3 cells was significantly reduced following exposure to apixaban (25 %), fondaparinux (12 %), enoxaparin (14 %) or tinzaparin (11 %). Exposure of BXPC3 to antithrombotic agents did not significantly modify the release of MP. Apixaban, fondaparinux, enoxaparin and tinzaparin decreased TG induced by the supernatant by 70 %, 30 %, 40 %, 90 % respectively. After exposure to BXPC3 cells the concentration of fondaparinux, enoxaparin and tinzaparin in the supernatant reduced by 27 %, 48 % and 26 % respectively. In contrast the concentration of apixaban did not significantly change.

Conclusion Antithrombotic agents do not alter cancer cells’ TF expression or procoagulant MP release, but inhibit the procoagulant potency of microenvironment. Nevertheless, a LMWHs and fondaparinux degradation occurs following two days of exposure to cancer cells. Antithrombotic agents reduced tumour cells’ viability and impaired mRNA expression of pro- and antiangiogenic factors.

Tab 1. Impact of antithrombotic agents on BXPC3 cells, on their MP, and on the capacity of trigger thrombin generation of their culture supernatants. Cells were treated with 2 Ul/ml of Fondaparinux/Apixaban, or 2 micro gram/ml of Tinzaparin/Enoxaparin for 48h. mRNA expression were normalized using the 2^(-delta(deltaCq)) method. * p < 0.05 as compare to Control experiment.

Publication History

Article published online:
18 June 2021