Background: Chronic myeloid leukemia (CML) is characterized by the spread of malignant cells that exhibit resistance to caspase-mediated cell death (apoptosis) and this mechanism is proposed to play an important role in myeloid cell growth. However, the extent to which caspase-mediated cell death plays a crucial role in the regulation of myelopoiesis remains controversial. Objectives & Proceedure: The objectives of this study were to examine whether or not caspase-mediated cell death-related proteins take part in the development of CML and to also to detect the relationship between Fas, p53 and caspase-mediated cell death protease activating factor (Apaf-1) in 5 patients with CML using the real-time quantitative polymerase chain reaction. We demonstratedthatp53 and Apaf-1 messenger ribonucleic acid (mRNA) expression was moderately elevated (up to 5 fold, p<0.05) in 4 out of 5 CML patients. One patient with a p53 point mutation, exhibited a far greater elevation of p53 mRNA expression throughout their blast crisis, but in contrast, displayed a significant reduction in levels of Apaf-1 mRNA and Fas mRNA. Conclusion: Our results show in-vivo linkages between Fas, p53 and Apaf-1 transcription parameters suggesting that the key genes involved in the caspase-mediated cell death might contribute to CML disease development.
Abstract:Introduction: Purifed CD34+ cells and cultured colony forming unit-granulocyte and macrophage (CFU-GM) from human bone marrow were utilized to examine the role of Fas/FasL and caspase-8interactionsin the regulation of myelopoiesis. Methods: Fas and FasL expression in CD34+ cells and in day 14 CFU-GM were measured by RT-PCR and immunofluorescence in each case. The functional tests for the CFU-GM were the standard settlement test and the proliferative limit of CFU-GM was measured by replating the essential cells and monitoring auxiliary colony development. Results: Treatment of CFU-GM with IETD significantly increased, the proliferative limit, while the Fas mAb resulted in decreased the Fas and FasL expression were measuredutilizing RT-PCR and immunofluorescence individually. Conclusion: Fas, FasL, and caspase attenuationplay important rolesin the control of myelopoiesis.
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