title: Regulation of tissue factor in NT2 germ cell tumor cells by cisplatin chemotherapy.
abstract: BACKGROUND
  Patients with germ cell tumors (GCTs) receiving cisplatin-based chemotherapy are at increased risk of thrombosis, but the underlying cellular and molecular mechanisms remain obscure.
  
  
  OBJECTIVE
  To study baseline tissue factor (TF) expression by GCT cell lines and its modulation by cisplatin treatment.
  
  
  METHODS
  TF expression was assessed by single-stage clotting and thrombin generation assay, flow cytometry, ELISA, and Western blot analysis. Cell cycle analysis and detection of phosphatidylserine (PS) membrane exposure were carried out by flow cytometry. TF mRNA was analyzed by quantitative RT-PCR.
  
  
  RESULTS
  Significant expression of TF-specific procoagulant activity (PCA) was detected on three non-seminoma (NT2, 2102Ep, NCCIT) and one seminoma cell line (TCam-2). Treatment with 0.4μM cisplatin (corresponding to the IC50) for 48hrs increased TF PCA on NT2 cells 3-fold, an effect that was largely independent of PS exposure and that could not be explained by translocation of active TF from intracellular storage pools. Cisplatin-induced TF PCA expression in NT2 cells did not occur before 12hrs, but was steady thereafter and accompanied by a 2-fold increase in total and surface-located TF antigen. Importantly, increased TF gene transcription or production and release of an intermediate factor were not involved in this process. Cell cycle analysis suggested that cisplatin-induced G2/M arrest resulted in an accumulation of procoagulant TF on the membrane surface of NT2 cells.
  
  
  CONCLUSIONS
  In addition to induction of apoptosis/necrosis with PS-mediated activation of preformed TF, cisplatin may alter the procoagulant phenotype of GCT cells through an increase in total cellular TF antigen.
authors.affiliation: II. Medizinische Klinik und Poliklinik, Onkologisches Zentrum - Universitäres Cancer Center Hamburg (UCCH), Universitätsklinikum Eppendorf, Martinistr. 52, D-20246 Hamburg, Germany.
authors.forename: Christine
authors.initials: C
authors.lastname: Jacobsen
authors.affiliation: II. Medizinische Klinik und Poliklinik, Onkologisches Zentrum - Universitäres Cancer Center Hamburg (UCCH), Universitätsklinikum Eppendorf, Martinistr. 52, D-20246 Hamburg, Germany.
authors.forename: Karin
authors.initials: K
authors.lastname: Oechsle
authors.affiliation: II. Medizinische Klinik und Poliklinik, Onkologisches Zentrum - Universitäres Cancer Center Hamburg (UCCH), Universitätsklinikum Eppendorf, Martinistr. 52, D-20246 Hamburg, Germany.
authors.forename: Jessica
authors.initials: J
authors.lastname: Hauschild
authors.affiliation: II. Medizinische Klinik und Poliklinik, Onkologisches Zentrum - Universitäres Cancer Center Hamburg (UCCH), Universitätsklinikum Eppendorf, Martinistr. 52, D-20246 Hamburg, Germany.
authors.forename: Gustav
authors.initials: G
authors.lastname: Steinemann
authors.affiliation: II. Medizinische Klinik und Poliklinik, Onkologisches Zentrum - Universitäres Cancer Center Hamburg (UCCH), Universitätsklinikum Eppendorf, Martinistr. 52, D-20246 Hamburg, Germany.
authors.forename: Brigitte
authors.initials: B
authors.lastname: Spath
authors.affiliation: II. Medizinische Klinik und Poliklinik, Onkologisches Zentrum - Universitäres Cancer Center Hamburg (UCCH), Universitätsklinikum Eppendorf, Martinistr. 52, D-20246 Hamburg, Germany.
authors.forename: Carsten
authors.initials: C
authors.lastname: Bokemeyer
authors.affiliation: Department of Immunology and Microbial Sciences, The Scripps Research Institute, 10550N Torrey Pines Rd, La Jolla, CA 92037, USA; Center for Thrombosis and Hemostasis, University Medical Center Mainz, Langenbeckstr. 1 D-55131 Mainz, Germany.
authors.forename: Wolfram
authors.initials: W
authors.lastname: Ruf
authors.affiliation: II. Medizinische Klinik und Poliklinik, Onkologisches Zentrum - Universitäres Cancer Center Hamburg (UCCH), Universitätsklinikum Eppendorf, Martinistr. 52, D-20246 Hamburg, Germany; Tumor and Breast Center ZeTuP, Rorschacher Str. 150, CH-9006 St. Gallen, Switzerland.
authors.forename: Friedemann
authors.initials: F
authors.lastname: Honecker
authors.affiliation: II. Medizinische Klinik und Poliklinik, Onkologisches Zentrum - Universitäres Cancer Center Hamburg (UCCH), Universitätsklinikum Eppendorf, Martinistr. 52, D-20246 Hamburg, Germany. Electronic address: langer@uke.de.
authors.forename: Florian
authors.initials: F
authors.lastname: Langer
mesh_terms: D000970:Antineoplastic Agents / Q000008:administration & dosage; D017209:Apoptosis / Q000187:drug effects; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D002945:Cisplatin / Q000008:administration & dosage*; D004305:Dose-Response Relationship, Drug; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D018922:HL-60 Cells; D006801:Humans; D009373:Neoplasms, Germ Cell and Embryonal / Q000188:drug therapy* / Q000378:metabolism* / Q000473:pathology; D013925:Thromboplastin / Q000378:metabolism*
chemical_list: D000970:Antineoplastic Agents; D013925:Thromboplastin; D002945:Cisplatin
keywords: Cancer; Chemotherapy; Phosphatidylserine; Thrombosis; Tissue factor