کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
3027085 | 1182938 | 2015 | 9 صفحه PDF | دانلود رایگان |

• Germ cell tumor (GCT) cell lines constitutively express tissue factor (TF)
• Cisplatin increases both TF procoagulant activity (PCA) and antigen in NT2 cells
• Induction of TF PCA by cisplatin is largely independent of phosphatidylserine
• Cisplatin-induced G2/M arrest is associated with TF accumulation in NT2 cells
BackgroundPatients with germ cell tumors (GCTs) receiving cisplatin-based chemotherapy are at increased risk of thrombosis, but the underlying cellular and molecular mechanisms remain obscure.ObjectiveTo study baseline tissue factor (TF) expression by GCT cell lines and its modulation by cisplatin treatment.MethodsTF 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.ResultsSignificant 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 0.4 μM cisplatin (corresponding to the IC50) for 48 hrs 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 12 hrs, 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.ConclusionsIn 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.
Journal: Thrombosis Research - Volume 136, Issue 3, September 2015, Pages 673–681