title: Progesterone utilizes distinct membrane pools of tissue factor to increase coagulation and invasion and these effects are inhibited by TFPI.
abstract: Tissue factor (TF) serving as the receptor for coagulation factor VII (FVII) initiates the extrinsic coagulation pathway. We previously demonstrated that progesterone increases TF, coagulation and invasion in breast cancer cell lines. Herein, we investigated if tissue factor pathway inhibitor (TFPI) could down-regulate progesterone-increased TF activity in these cells. Classically, TFPI redistributes TF-FVII-FX-TFPI in an inactive quaternary complex to membrane associated lipid raft regions. Herein, we demonstrate that TF increased by progesterone is localized to the heavy membrane fraction, despite progesterone-increased coagulation originating almost exclusively from lipid raft domains, where TF levels are extremely low. The progesterone increase in coagulation is not a rapid effect, but is progesterone receptor (PR) dependent and requires protein synthesis. Although a partial relocalization of TF occurs, TFPI does not require the redistribution to lipid rafts to inhibit coagulation or invasion. Inhibition by TFPI and anti-TF antibodies in lipid raft membrane fractions confirmed the dependence on TF for progesterone-mediated coagulation. Through the use of pathway inhibitors, we further demonstrate that the TF up-regulated by progesterone is not coupled to the progesterone increase in TF-mediated coagulation. However, the progesterone up-regulated TF protein may be involved in progesterone-mediated breast cancer cell invasion, which TFPI also inhibits.
authors.affiliation: Departamento de Fisiología, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, Alameda 340, Santiago, Chile.
authors.forename: Soledad
authors.initials: S
authors.lastname: Henriquez
authors.forename: Claudia
authors.initials: C
authors.lastname: Calderon
authors.forename: Marisol
authors.initials: M
authors.lastname: Quezada
authors.forename: Bárbara
authors.initials: B
authors.lastname: Oliva
authors.forename: Maria Loreto
authors.initials: ML
authors.lastname: Bravo
authors.forename: Evelyn
authors.initials: E
authors.lastname: Aranda
authors.forename: Sumie
authors.initials: S
authors.lastname: Kato
authors.forename: Mauricio A
authors.initials: MA
authors.lastname: Cuello
authors.forename: Jorge
authors.initials: J
authors.lastname: Gutiérrez
authors.forename: Andrew F G
authors.initials: AF
authors.lastname: Quest
authors.forename: Gareth I
authors.initials: GI
authors.lastname: Owen
mesh_terms: D001777:Blood Coagulation; D001943:Breast Neoplasms / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism*; D015942:Factor VIIa / Q000378:metabolism; D005170:Factor X / Q000378:metabolism; D005260:Female; D006801:Humans; D008074:Lipoproteins / Q000378:metabolism*; D021962:Membrane Microdomains / Q000378:metabolism; D009361:Neoplasm Invasiveness; D011374:Progesterone / Q000378:metabolism*; D021381:Protein Transport; D011980:Receptors, Progesterone / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D013925:Thromboplastin / Q000378:metabolism*; D013997:Time Factors; D015854:Up-Regulation
chemical_list: D008074:Lipoproteins; D011980:Receptors, Progesterone; D011994:Recombinant Proteins; C051928:lipoprotein-associated coagulation inhibitor; D011374:Progesterone; D005170:Factor X; D013925:Thromboplastin; D015942:Factor VIIa