Aggressiveness of breast cancer cells is related with tissue factor expression and procoagulant activity.

Objective The procoagulant potency of cancer cells may vary according to the histological type and their aggressiveness. We identified the procoagulant « fingerprint » of breast cancer cells lines of various degrees of aggressiveness.

Material and Methods Breast adenocarcinoma cell lines MCF7, MCF7-sh-WISP2, BT-20 and MDAMB-321 were used. Flow cytometry and western blot analysis for TF expression were performed using an anti-human TF murine IgG1 monoclonal antibody. Cancer cells were added in platelet poor plasma (PPP) from healthy volunteers and thrombin generation (TG) was assessed using Calibrated Automated Thrombogram.

Results Incubation of MCF7, MCF7-sh-WISP2, MDAMB-231, BT-20 and HUVEC cells with PPP resulted in acceleration of the initiation phase of thrombin generation. BT-20 manifested higher procoagulant potential than MCF7-sh-WISP2 than MCF7wt than MDAMB-231;

Flow cytometry analysis of TF expression showed a significantly higher expression on the membrane of MCF7-sh-WISP2 and BT-20. Western blot analysis showed that TF was present in eluted proteins after immunoprecipitation of cell lysates with anti-TF and in cell lysates. Two different major bands were observed at levels of 47 kDa and 25 kDa. In MCF7 a significant 47 KDa band was observed though its intensity was lower compared to the corresponding band of the lysate from MCF7-sh-WISP2; MDAMB-231; BT-20 cells.

Conclusion We demonstrate that procoagulant phenotype of pancreatic and breast cancer, is related to the expression of functional TF. The procoagulant fingerprint of breast cancer cells varies in function of the degree of aggressiveness. MCF7-ShWisp2 and BT-20 were the more aggressive cells and present higher procoagulant potential which is correlated with TF expression on the cell membrane. The present experimental model will allow the characterization of the procoagulant fingerprint of cell lines from the same or different histological types of cancer. It will also allow toevaluate the efficiency of antithrombotic agents to downregulate hypercoagulability induced by cancer cells.

Publication History

Article published online:
18 June 2021