Activated protein C upregulates ovarian cancer cell migration and promotes unclottability of the cancer cell microenvironment

Althawadi,Hamda; Alfarsi,Halema; Besbes,Samaher; Mirshahi,Shahsoltan; Ducros,Elodie; Rafii,Arash; Pocard,Marc; Therwath,Amu; Soria,Jeannette; Mirshahi,Massoud

doi:10.3892/or.2015.4061

Activated protein C upregulates ovarian cancer cell migration and promotes unclottability of the cancer cell microenvironment

Authors:
- Hamda Althawadi
- Halema Alfarsi
- Samaher Besbes
- Shahsoltan Mirshahi
- Elodie Ducros
- Arash Rafii
- Marc Pocard
- Amu Therwath
- Jeannette Soria
- Massoud Mirshahi
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Affiliations: UMR, Paris Diderot, Paris 7 University, Lariboisiere Hospital, INSERM U965, Paris, France, Qatar Foundation, Weill Cornell Medical College in Qatar, Doha, Qatar
Published online on: June 15, 2015 https://doi.org/10.3892/or.2015.4061
Pages: 603-609
Copyright: © Althawadi et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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Abstract

The objective of this study was to evaluate the role of activated protein C (aPC), known to be a physiological anticoagulant, in ovarian cancer cell activation as well as in loss of clotting of cancer ascitic fluid. The effect of aPC on an ovarian cancer cell line (OVCAR-3) was tested in regards to i) cell migration and adhesion with the use of adhesion and wound healing assays as well as a droplet test; ii) protein phosphorylation, evaluated by cyto-ELISA; iii) cell cycle modification assessed by flow cytometric DNA quantification; and iv) anticoagulant activity evaluated by the prolongation of partial thromboplastin time (aPTT) of normal plasma in the presence or absence of aPC‑treated ovarian cancer cells. In addition, the soluble endothelial protein C receptor (sEPCR) was quantified by ELISA in ascitic fluid of patients with ovarian cancer. Our results showed that in the OVCAR-3 aPC-induced cells i) an increase in cell migration was noted, which was inhibited when anti-endothelial protein C receptor (EPCR) was added to the culture medium and which may act via MEK-ERK and Rho-GTPase pathways; ii) an increase in threonine, and to a lesser extent tyrosine phosphorylation; iii) cell cycle activation (G1 to S/G2); and iv) a 2-3-fold prolongation of aPTT of normal plasma. In the peritoneal fluid, the sEPCR concentration was 71±23 ng/ml. In conclusion, free aPC binds to membrane EPCR in ovarian cancer cells and induces cell migration via MEK-ERK and Rho-GTPase pathways. This binding could also explain the loss of clotting of peritoneal fluids.

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