Differentiating effect of thalidomide and GM-CSF combination on HL-60 acute promyelocytic leukemia cells

Aim: To investigate whether granulocyte-macrophage colony-stimulating factor (GM-CSF) with or without thalidomide can induce apoptosis and differentiation of HL-60 acute promyelocytic leukemia cell line in vitro. Methods: Effect of GM-CSF and thalidomide on proliferation of HL-60 cells was evaluated by MTT assay, cell cycle analysis was performed by propidium iodide staining approach and flow cytometry, and apoptosis rate was analyzed using FITC-conjugated annexin-V and FACScan flow cytometry. Results: The study revealed that thalidomide alone at high concentrations inhibited HL-60 cell growth and induced apoptosis. Three days treatment of low-dose thalidomide in combination with GM-CSF induced marked terminal differentiation of HL-60 cells, as it was assessed by increased expression of differentiation antigens on cell surface. Conclusion: Treatment of HL-60 cells by low concentration of thalidomide combined with GM-CSF induced terminal differentiation of HL60 cells in vitro, which may be advantageous for the elaboration of novel therapeutic regimens in patients with differentiation-inducible leukemias.Цель: изучить эффект гранулоцитарно-макрофагального колониестимулирующего фактора (ГМ-КСФ) в сочетании с талидомидом на индукцию апоптоза и дифференцировку клеток острого промиелоцитарного лейкоза линии HL-60 in vitro. Методы: для оценки пролиферации и жизнеспособности клеток HL-60 применяли MTT анализ, для изучения клеточного цикла — окраску пропидиум бромидом и проточную цитометрию. Для оценки апоптоза клетки линии HL-60 обрабатывали талидомидом, ГМ-КСФ, и совместно талидомидом и ГМ-КСФ в течении 48 ч, и затем метили анексином, конъюгированным с FITC, и анализировали с помощью проточной цитометрии. Результаты: талидомид в высоких концентрациях ингибирует пролиферацию клеток HL-60 и вызывает апоптоз. В сочетании с ГМ-КСФ в течение 3 дней талидомид в низкой концентрации индуцировал терминальную дифференцировку клеток HL-60, о чем свидетельствовало появление экспрессии дифференцировочных антигенов на поверхности клеток. Выводы: применение талидомида в низкой концентрации в сочетании с ГМ-КСФ вызывает терминальную дифференцировку клеток HL-60

Anaerobik Glikoliz HL-60 Akut Promyelositik Lösemi Hücrelerinde Enerjinin Oluşumu için Temel Yoldur

Aim In physiological conditions, normal cells use mainly the glycolytic aerobic pathway to provide energy. However, most cancer cells utilize anaerobic glycolytic way for energy generation. Aim of this study was to investigate the carbohydrate metabolic pathways of HL-60 acute promyelocytic leukemia cells for energy production. Material and Methods Leukemia cells as well as normal leukocytes were incubated with radiolabelled glucose in aerobic and anaerobic conditions and glycogen consumptions and the ratios of radiolabelled glucose catabolized into CO2 or lactate, that is, the rates of aerobic or anaerobic glycolysis, were determined. Results The glycogen consumption was significantly higher in aerobic leukemia cell culture than normal leukocyte culture (p<0.01). The rate of anaerobic glycolysis was 93.8% in leukemia cells in aerobic conditions and it increased to 96.6% while utilization of glycogen increased by 7.31% in anaerobic conditions. Conclusion In conclusion, principally anaerobic glycolysis is effective for energy generation in HL-60 promyelocytic leukemia cells. This result may be important for the development of new therapeutic approaches in the treatment of promyelocytic leukemia, requiring further comprehensive studies

Alteration of tumor glucose metabolism after radiotherapy in MCF-7 breast cancer cell lines

Cancer cells utilize anaerobic glycolytic way to compensate their faster metabolism when compared to normal cells. The purpose of this study is to investigate the effect of radiation on tumor metabolism. MCF-7 breast cancer cell lines were divided into 4 groups, including 2 control groups and aerobic and anaerobic study groups (were irradiated 600 cGy by Co-60 teletherapy unit), incubated with radiolabelled glucose for 4 hours. One control group was for aerobic, and the other was for anaerobic group after KCN addition. Radiolabelled CO2 produced by the cells was isolated and collected in specially designed simulation vials. In supernatant the measurements of other end-products of carbohydrate catabolism including lactate, pyruvate, acetate were performed on a liquid scintillation analyzer after they were collected via anion-exchange chromatography. Finally glucose in supernatant was measured enzymatically by glucose oxidase method. Glycogen consumption and lactate production were significantly higher in anaerobic and radiation groups (p<0.01). Whereas CO2 production was significantly higher in control group (p<0.01). Taken all results together radiation lead tumor cells more anaerobic glycolysis with high glycogen consumption, high lactate production and low CO2 production. Radiation itself has led tumor cells to produce energy by anaerobic glycolysis, meaning radiation exposed cells become more hypoxic

Protein profiling of anastomosed facial nerve treated with mesenchymal stromal cells

PubMed: 22268520Background aims. The types of proteins released from mesenchymal stromal cells (MSC) are still unclear. Our aim was to compare apoptosis scores and the expression of myelin-associated glycoprotein (MAG), myelin basic protein (MBP), neural cell adhesion molecule (NCAM)-1,matrix metalloproteinase (MMP)-1A, tissue inhibitor of metalloproteinase (TIMP)-1, TIMP-1/MMP-1A ratio, nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), ciliary neurotrophic factor (CNTF), neurotrophin (NT)-3, NT-4, glial cell-derived neurotropic factor (GDNF), leukemia inhibitory factor (LIF), basic fibroblast growth factor (FGF)-2, insulin-like growth factor (IGF)-1, platelet-derived growth factor (PDGF)-? and transforming growth factor (TGF)-?1 in anastomosed facial nerves that had been treated with or without MSC. Methods. In seven rats, the buccal branch of the right facial nerve was transected, anastomosed and treated with MSC (anastomosed + MSC group). The left buccal branch was anastomosed only (anastomosed-only group). The left mandibular branch served as an intact nerve group. On days 1820, the distal segments of the branches were examined in terms of expression of the mentioned proteins and apoptosis scores using polymerase chain reaction (PCR) and terminal deoxynucleotidyl transferase-mediated digoxigenin-UTP nick end labeling (TUNEL) assays. Results. MSC application significantly increased CNTF, PDGF-?, LIF, TGF-?1, BDNF and NT-3 expression (P 0.05). Changes in other proteins and apoptosis scores were not significant. Conclusions. These results suggest that MSC increases expression of CNTF, PDGF-?, LIF,TGF-?1, BDNF and NT-3. MAG, NCAM-1, MMP-1A and FGF-2 expressions were slightly changed in this stage of nerve regeneration. The comparison of apoptotic activity was not conclusive. Overall, it appears that MSC might have differential effects on the mentioned tissue-related proteins and trophic/growth factors. © 2012 Informa Healthcare.The authors should thank Meral SARPER, MSc, and Pinar ELCI, MSc, from the Cancer Research Center and Medical Research Center of Gulhane Military Medical Academy for preparing stem cells. The authors are grateful to Tayfun Ide, Veterinary Surgeon, Manager of the Surgical Research Section, for providing maintenance of the animals. Conflict of interest: The authors have no conflict of interest. The authors are grateful to the Research and Development Center/Gulhane Military Medical Academy (Ankara/Turkiye) that funded the research

Current practice of autologous hematopoietic progenitor cell mobilization in adult patients with multiple myeloma and lymphoma: The results of a survey from Turkish hematology research and education group (ThREG)

Autologous hematopoietic cell transplantation (AHCT) is an established treatment option for adult patients presenting with multiple myeloma (MM), Hodgkin lymphoma (HL) and various subtypes of non-Hodgkin lymphoma (NHL) in upfront and/or relapsed/refractory disease settings. Although there are recently published consensus guidelines addressing critical issues regarding autologous hematopoietic progenitor cell mobilization (HPCM), mobilization strategies of transplant centers show high variability in terms of routine practice. In order to understand the current institutional policies regarding HPCM in Turkey and to obtain the required basic data for preparation of a national positional statement on this issue, Turkish Hematology Research and Education Group (ThREG) conducted a web-based HPCM survey. The survey was designed to include multiple-choice questions regarding institutional practice of HPCM in adults presenting MM, HL, and NHL. The representatives of 27 adult HCT centers participated to the study. Here we report the results of this survey shedding light on the real-world experience in Turkey in terms of autologous HPCM mobilization strategies in patients presenting with MM and lymphoma. © 2017 Elsevier Lt