Abstract
Aim:
The differentiation capability of guanosine 5′-triphosphate (GTP) was studied using U937 and KG1 cells.
Methods:
Cell cycle was analyzed by PI staining using flow cytometry. Apoptosis was measured by Annexin-V-FITC/PI double staining using flow cytometry. Differentiation was observed by morphological criteria, Wright-Giemsa staining and expression of cell surface markers CD11b and CD14.
Results:
Variable GTP concentrations (25–200 μmol/L) at short treatment times (up to 24 h) showed significant anti-proliferative activities among both cell types. However, longer treatment times (up to 72 h) were required to trigger apoptosis. Cell-cycle analyses of the GTP-treated cells indicated an increase in S-phase population by 48 h followed by the appearance of a sub-G1 peak after 72 h of treatment. The effects of GTP on U937 and KG1 cells were accompanied with differentiation toward monocyte/macrophage lineage. This was evidenced by a sharp increase in the extent of CD11b and CD14 expression after 24 h of exposure to GTP. The viability of both cell types did not significantly change during the first 24 h. However, at longer treatment times (72–96 h), dramatic decreases in both the extent of CD 14 expression and the cell viabilities were observed. Simultaneous measurement of apoptosis and CD14 expression in GTP-treated U937 cells indicated that cells with lower CD 14 content underwent more apoptosis.
Conclusion:
These finding may pave the way for further pharmaceutical evaluation of GTP as a suitable differentiating agent for acute myeloblastic leukemia (AML) therapy.
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References
Coco FL, Nervi C, Avvisati G, Mandelli F . Acute promyelocytic leukemia: a curable disease. Leukemia 1998; 12: 1866–80.
Stegmaier K, Corsello SM, Ross KN, Wong JS, DeAngelo DJ, Golu TR . Gefitinib induces myeloid differentiation of acute myeloid leukemia. Blood 2005; 106: 2841–8.
Wang ZY . Ham-Wasserman lecture: treatment of acute leukemia by inducing differentiation therapy and apoptosis. Hematology (Am Soc Hematol Edu Program) 2003; 1–13.
Spira AL, Carducci MA . Differentiation therapy. Curr Opin Pharmacol 2003; 3: 338–436.
Bruserud O, Gjertsen BT . New strategies for the treatment of acute myelogenous leukemia: differentiation induction - present use and future possibilities. Stem Cells 2000; 18: 157–65.
Sachs L . The control of hematopoiesis and leukemia: from basic biology to the clinic. Proc Natl Acad Sci USA 1996; 93: 4742–9.
Sundstron C, Nillsson K . Establishment and characterization of a human histiocytic lymphoma cell line (U-937). Int J Cancer 1976; 17: 565–72.
Koeffler HP, Golde DW . Human myeloid leukemia cell lines: a review. Blood 1980; 56: 344–50.
Rizzo MG, Zepparoni A, Cristofanelli B, Scardigli R, Crescenzi M, Blandio G, et al. Wt-p53 action in human leukemia cell lines corresponding to different stages of differentiation. Br J Cancer 1998; 77: 1429–38.
Koeffler HP, Billing R, Sparkes RS, Golde DW . An undifferentiated variant derived from the human acute myelogenous leukemia cell line (KG-1). Blood 1980; 56: 265–73.
Ahmed N, Laverick L, Sammons J, Baumforth KRN, Hassan HT . Effect of all-trans retinoic acid on chemotherapy induced apoptosis and down-regulation of Bc1-2 in human myeloid leukaemia CD34 positive cells. Leukemia Res 1999; 23: 741–9.
Wang H, Zheng X, Behm FG, Ratnam M . Differentiation-independent retinoid induction of folate receptor type beta, a potential tumor target in myeloid leukemia. Blood 2000; 96: 3529–36.
Chow SC, Kass GE, Orrenius S . Purines and their role in apoptosis. Neuropharmacology 1997; 36: 1149–56.
Lemoli RM, Ferrari D, Fogli M, Rossi L, Pizzirani C, Forchap S, et al. Extracellular nucleotides are potent stimulators of human hematopoietic stem cells in vitro and in vivo. Blood 2004; 104: 1662–70.
Rathbone MP, Middlemiss PJ, Gysbers JW, Andrew C, Herman MA, Reed JK, et al. Trophic effects of purines in neurons and glial cells. Prog Neurobiol 1999; 59: 663–90.
Schneider C, Wiendl H, Ogilvie A . Biphasic cytotoxic metabolism of extracellular ATP on U-937 human histiocytic leukemia cells: involvement of adenosine generation. Biochim Biophys Acta 2001; 1538: 190–205.
Rapaport E . Treatment of human tumor cells with ADP or ATP yields arrest of growth in the S phase of the cell cycle. J Cell Physiol 1983; 114: 279–83.
Di Iorio, P, Ballerini P, Teraversa U, Nicoletti F, Alimonti I, Kleywegt S, et al. The antiapoptotic effect of guanosine is mediated by activation of the PI3-kinase/AKT/PBK pathway in culture rat astrocytes. Neuroscience 2004; 46: 356–68.
Giotta GJ, Smith JR, Nicolson GL . Guanosine 5′-triphosphate inhibits growth and stimulates differentiated functions in B16 melanoma cells. Exp Cell Res 1978; 15: 385–93.
Osti F, Corradini FG, Hanau S, Matteuzzi M, Gambari R . Human leukemia K562 cells: induction of erythriod differentiation by guanine, guanosine and guanine nucleotides. Haematologica 1997; 82: 395–401.
Morceau F, Dupont C, Palissot V, BordChieche P, Trentesax C, Dicato M, et al. GTP-mediated differentiation of the human K562 cell line: transient overexpression of GATA-1 and stabilization of the α-globin. Leukemia 2000; 14: 1589–97.
Nicoletti I, Migliorati G, Pagliacci MC, Grignani F, Riccardi C . A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods 1991; 139: 271–9.
Yazdanparast R, Moosavi MA, Mahdavi M, Sanati MH . 3-Hydrogenkwadaphnin from dendrostellera lessertii induces differentiation and apoptosis in HL-60 cells. Planta Med 2005; 71: 1112–7.
Gockel HR, Lugering A, Heidemann J, Schmidt M, Domschke W, Kucharzik T, et al. Thalidomide induces apoptosis in human monocytes by using a cytochrome c-dependent pathway. J Immunol 2004; 172: 5103–9.
Hassan HT . Ajoene (natural garlic compound): a new anti-leukaemia agent for AML therapy. Leukemia Res 2004; 28: 667–71.
Hass R, Bartels H, Topley N, Hadam M, Köhler L, Goppelt-Strube M, et al. TPA-induced differentiation and adhesion of U937 cells: changes in ultrastructure, cytoskeletal organization and expression of cell surface antigens. Eur J Cell Biol 1989; 48: 282–93.
Prudovsky I, Popov K, Akimov S, Servo S, Zelenin A, Meinhardt G, et al. Antisense CD11b integrin inhibits the development of a differentiated monocyte/macrophage phenotype in human leukemia cells. Eur J Cell Biol 2002; 81: 36–42.
Bernhard D, Tinhofer I, Toko M, Hubi H, Ausserlechner MJ, Gril R, et al. Resveratrol causes arrest in the S-phase prior to Fas-independent apoptosis in CEM-C7H2 acute leukemia cells. Cell Death Differ 2000; 7: 834–42.
Mlejnek P, Kozubek S . Adenine-induced arrest of mammalian cells in early S-phase is related to the prevention of DNA synthesis inhibition caused by gamma-irradiation. Int J Radiol Biol 1997; 71: 505–13.
Carvalhal AV, Santos SS, Calado J, Haury M, Carrondo MJ . Cell growth arrest by nucleotides, nucleosides and bases as a tool for improved production of recombinant proteins. Biothechnol Prog 2003; 19: 69–83.
Heidenreich S, Schmidt M, August C, Cullen P, Rademaekers AH, Pauels G . Regulation of human monocyte apoptosis by the CD14 molecule. J Immunol 1997; 159: 3178–88.
Zhang DE, Hetherington CJ, Gonzalez DA, Chen HM, Tenen DG . Regulation of CD 14 expression during monocytic differentiation induced with 1α, 25-dihydroxyvitamin D3. J Immunol 1994; 153: 3276–84.
Kanatani Y, Kasukabe T, Okabe-Kado J, Yamamoto-Yamaguchi Y, Nagata N, Motoyoshi K, et al. Role of CD14 expression in the differentiation-apoptosis switch in human monocytic leukemia cells treated with 1α, 25-dihydroxyvitamin D3 or dexam-ethasone in the presence of transforming growth factor β. Cell Growth Differ 1999; 10: 705–12.
Brackman D, Lund-Johansen F, Aarskog D . Expression of leukocyte differentiation antigens during the differentiation of HL-60 cells induced by 1α, 25-dihydroxyvitamin D3: comparison with the maturation of normal monocytic and granulocytic bone marrow cells. J Leukoc Biol 1995; 58: 547–55.
Guzman ML, Jordan CT . Considerations for targeting malignant stem cells in leukemia. Cancer Control 2004; 11: 97–104.
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Project supported by the research council of the University of Tehran and the National Research Center for Genetic Engineering and Biotechnology, Tehran, Iran.
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Yazdanparast, R., Moosavi, M., Mahdavi, M. et al. Guanosine 5′-triphosphate induces differentiation-dependent apoptosis in human leukemia U937 and KG1 cells. Acta Pharmacol Sin 27, 1175–1184 (2006). https://doi.org/10.1111/j.1745-7254.2006.00364.x
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DOI: https://doi.org/10.1111/j.1745-7254.2006.00364.x
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