title: Activated protein C upregulates ovarian cancer cell migration and promotes unclottability of the cancer cell microenvironment.
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.
authors.affiliation: UMR, Paris Diderot, Paris 7 University, Lariboisiere Hospital, INSERM U965, Paris, France.
authors.forename: Hamda
authors.initials: H
authors.lastname: Althawadi
authors.affiliation: UMR, Paris Diderot, Paris 7 University, Lariboisiere Hospital, INSERM U965, Paris, France.
authors.forename: Halema
authors.initials: H
authors.lastname: Alfarsi
authors.affiliation: UMR, Paris Diderot, Paris 7 University, Lariboisiere Hospital, INSERM U965, Paris, France.
authors.forename: Samaher
authors.initials: S
authors.lastname: Besbes
authors.affiliation: UMR, Paris Diderot, Paris 7 University, Lariboisiere Hospital, INSERM U965, Paris, France.
authors.forename: Shahsoltan
authors.initials: S
authors.lastname: Mirshahi
authors.affiliation: UMR, Paris Diderot, Paris 7 University, Lariboisiere Hospital, INSERM U965, Paris, France.
authors.forename: Elodie
authors.initials: E
authors.lastname: Ducros
authors.affiliation: Qatar Foundation, Weill Cornell Medical College in Qatar, Doha, Qatar.
authors.forename: Arash
authors.initials: A
authors.lastname: Rafii
authors.affiliation: UMR, Paris Diderot, Paris 7 University, Lariboisiere Hospital, INSERM U965, Paris, France.
authors.forename: Marc
authors.initials: M
authors.lastname: Pocard
authors.affiliation: UMR, Paris Diderot, Paris 7 University, Lariboisiere Hospital, INSERM U965, Paris, France.
authors.forename: Amu
authors.initials: A
authors.lastname: Therwath
authors.affiliation: UMR, Paris Diderot, Paris 7 University, Lariboisiere Hospital, INSERM U965, Paris, France.
authors.forename: Jeannette
authors.initials: J
authors.lastname: Soria
authors.affiliation: UMR, Paris Diderot, Paris 7 University, Lariboisiere Hospital, INSERM U965, Paris, France.
authors.forename: Massoud
authors.initials: M
authors.lastname: Mirshahi
mesh_terms: D000925:Anticoagulants / Q000378:metabolism*; D015703:Antigens, CD / Q000378:metabolism*; D001202:Ascitic Fluid / Q000378:metabolism* / Q000473:pathology; D002448:Cell Adhesion / Q000187:drug effects; D045744:Cell Line, Tumor; D002465:Cell Movement; D000074283:Endothelial Protein C Receptor; D005260:Female; D005337:Fibrin / Q000378:metabolism; D006801:Humans; D010051:Ovarian Neoplasms / Q000378:metabolism / Q000473:pathology*; D010766:Phosphorylation; D011486:Protein C / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000378:metabolism*; D015398:Signal Transduction; D059016:Tumor Microenvironment; D015854:Up-Regulation
chemical_list: D000925:Anticoagulants; D015703:Antigens, CD; D000074283:Endothelial Protein C Receptor; C478370:PROCR protein, human; D011486:Protein C; D011956:Receptors, Cell Surface; D005337:Fibrin