Effect of ionizing radiation on cellular procoagulability and co-ordinated gene alterations

From the Department of Cardiology and Pneumology, Charité-Universitätsmedizin Berlin, Campus Benjamin Franklin, Berlin, Germany (PG-L, KP, Q-VT, BS, FW, SA, TW, WP, H-PS, UR); Center for Cardiovascular Research, Charité-Universitätsmedizin Berlin, Campus Mitte, Berlin, Germany (HW); Department of Pathology, Charité-Universitätsmedizin Berlin, Campus Benjamin Franklin, Berlin, Germany (MH, DL) Correspondence: Ursula Rauch, MD, Medical Clinic II, Charité-Universitätsmedizin Berlin Campus Benjamin Franklin, Hindenburgdamm 30, 12200 Berlin, Germany. E-mail: ursula.rauch{at}charite.de Background and Objectives: Ionizing radiation (IR) is associated with thrombotic vascular occlusion predicting a poor clinical outcome. Our study examined whether IR induced tissue factor (TF) expression and procoagulability. We further investigated coordinated gene alterations associated with TF upregulation in the myelomonocytic leukemia THP-1 cells. Design and Methods: TF expression was determined by quantitative Reverse Transcriptase (TaqMan ® ) PCR, TF ELISA and TF activity by a two stage chromogenic assay in the time course of days 1, 3, 7, 10, and 17 post IR. To detect IR-induced alterations in gene expression, Affymetrix HG U133 Plus 2.0 microarrays were used. Results: IR induced a significant increase in TF/GAPDH mRNA ratios and cellular TF protein on days 3 and 7 post IR (20 Gy [ p 0.01] and 40 Gy [ p 0.01 vs. control]), suggesting a late and persistent induction of TF. An increase in cellular TF activity was already found 1 day post IR (20 Gy and 40 Gy [ p 0.001] vs. control respectively), suggesting IR immediately alters the cellular thrombogenicity. TF upregulation post IR was confirmed in PBMNCs. Gene expression profiling showed IR increased the expression of inflammatory and apoptosis-related pathways known to be involved in the regulation of TF expression. Interpretation and Conclusions: TF upregulation together with inflammation and apoptosis may increase the thrombogenicity of tissues. The demonstrated upregulation of TF might play a pivotal role in radiation associated thrombosis. Key words: gene expression profiles, ionizing radiation, myelomonocytic cell line THP-1, procoagulability, tissue factor.