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Angiogensis plays the crucial role in growth and dissemination of neoplastic diseases, both for solid tumors and hematopoietic malignancies. Development of the abundant neoplastic vasculature results from an imbalance between pro- and antiangiogenic regulatory mechanisms. The investigation was focused on expression of the main regulatory angiogeneic factors in different phases of chronic myeloid leukemia (CML) and on influence of leukemic cells on the human umbilical vein endothelial cells proliferation. The groups of 29 patients with CML and of 14 healthy controls were enrolled to the study. The expression of vascular endothelial growth factor (VEGF), basic fibroblast growth factor, interleukin-8, angiopoetin-1, platelet factor-4, extracellular matrix metalloproteinases (MMP) -2 and -9 as well as tissue inhibitors of metalloproteinases was determined in peripheral blood and bone marrow mononuclear cells with the use of quantitative real-time PCR method and additionally the concentration of VEGF with the flow cytofluorometry. We evaluated the phosphorylation of endothelial mitogen activated protein kinase in human umbilical vein endothelial cells after incubation in CML cells conditioned media, applying Western blot technique. We determined an influence of the leukemic cells on the endothelial cells proliferation with the colorimetric metabolic MTT test. We showed, that in peripheral blood and bone marrow mononuclear cells in CML patients most of the studied factors were increased at the time of CML diagnosis and became lower in remission. In newly diagnosed CML patients an expression of VEGF, MMP-2 and MMP-9 was particularly elevated. In remission, the levels of VEGF and metalloproteinases, specifically MMP-9, were decreased. If failed to achieve remission, the patients presented the elevated expression of most of the investigated angiogenic factors. In the acceleration or blast crisis phase angiopoetin, VEGF and MMP-2 levels were particularly high. We noticed the markedly enhanced human umbilical vein endothelium proliferation after an incubation in CML cells conditioned media, both in the test of mitogen activated protein kinase phosphorylation in endothelial cells and in the metabolic test for the proliferation intensity of endothelium. The differences of an angiogeneic potential found between clinical phases of CML, and the ability of leukemic cells to stimulate endothelial proliferation, point at the significance of neovascularisation in CML pathogenesis. Further studies are necessary to delineate the possibility of the use of angiogenic inhibitors in the treatment of this malignancy.

Unlabelled: Chronic myeloid leukemia (CML) is one of the chronic myeloproliferative disorders. Haemostatic disturbances and infections are the main causes of death in CML., Material and Methods: We investigated 22 patients with CML aged 38-55 years (median 45.0). Twenty nine healthy controls were sex and age-matched. Patients underwent following examination: whole blood count, blood smear, platelets count, prothrombin time (PT), activated partial thromboplastin time (aPTT), euglobulin lysis time (ELT), fibrin degradation products (FDP), thrombin-antithrombin complexes (TAT), plasmin-alpha2-antiplasmin complexes (PAP), antigen and urokinase plasminogen activators (t-PA:Ag, u-PA:Ag), antigen of tissue plasminogen activator inhibitors type 1 and 2 (PAI-1:Ag, PAI-2:Ag), fibrinogen concentrations, antitrombin (AT) and alpha2-antiplasmin (alpha2-AP) activity., Results: TAT concentration (35.46 ng/ml) was significantly higher in examined group than in controls (3.41 ng/ml). Significantly higher fibrinogen concentration (3.31 g/l) and elevated platelet count (611.0 G/1) was observed in patients with CML. We also showed significantly higher concentrations of u-PA:Ag (0.67 ng/ml), PAI-1:Ag (34.8 ng/ml), PAP complexes (473.10 ng/ml) and FDP (17.10 microg/ml) in patients with CML., Conclusion: High TAT, fibrinogen concentrations and elevated platelet count in patients with CML are the evidences of an activation of coagulation. On the other hand fibrinolysis activation is proved by higher concentrations of u-PA:Ag, PAI-1:Ag, complexes PAP and FDP.

We examined the hybrid bcr/abl mRNA present in 59 patients with chronic myeloid leukemia, using the reverse transcription method and polymerase chain reaction. Bcr/abl gene was found in 98% of patients. 60% of patients had b3a2 type of translocation, 40% type b2a2. Patients with b3a2 type had higher platelet count at diagnosis presentation than patients with b2a2. Other hematological data were similar in both groups.