Abstract
The antileukemic activity of cytotoxic drugs is increasingly thought to be the result of induction of apoptosis. Several proto-oncogenes have been related to the regulation of this process. In this study we evaluated the relation between bcl-2 expression, spontaneous and dexamethasone (DXM) induced apoptosis, and in vitro resistance to DXM, prednisolone (PRD) and cytarabine (ARA) determined using the total cell kill colorimetric methyl-thiazol-tetrazolium salt (MTT) assay, in childhood acute lymphoblastic leukemia (ALL). Drug resistance was expressed as the LC50 value, the drug concentration lethal to 50% of the cells. Fourty-six samples taken at initial diagnosis (iALL) and 31 samples taken at relapse (rALL) were incubated in culture medium, with and without DXM. Bcl-2 expression and apoptosis were measured flowcytometrically, the latter using DNA histogram analysis. Bcl-2 expression was 1.4 fold higher in rALL than in iALL (p=0.008). Both spontaneous and DXM induced apoptosis increased significantly from 0 to 48 hours (in up to 71%, 81% of the cells respectively). Bcl-2 expression was inversely correlated with the extent of spontaneous apoptosis after 24 hours in iALL (r=-0.40, p=0.05). Relapsed samples, but not samples obtained at presentation, expressing high levels of bcl-2 displayed increased resistance to drug induced apoptosis (r=-0.63, p= 0.02). In iALL high bcl-2 expression appeared to be related to low LC50 values of ARA. No correlations were found for DXM or PRD. In conclusion, DXM excerts its cytotoxic effect at least partly by means of induction of apoptosis. Bcl-2 inhibits drug induced apoptosis in rALL. However in iALL bcl-2 expression is not associated with increased in vitro drug resistance, nor with increased resistance to drug induced apoptosis.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Yunis JJ, Oken MM, Kaplan ME, Ensrud KM, Howe RR, Theologides A. Distinctive chromosomal abnormalities in histologic subtypes of non-Hodgkin’s lymphoma. New England Journal of Medicine 1982, 307:1231–1236
Tsujimoto Y. Overexpression of the human bcl-2 gene product results in growth enhancement of Ebstein-Barr virus immortalized B cells. Proc Nat Acad Sci USA 1984, 86:1958–1962
Cleary ML, Smith SD, Sklar J. Cloning and structural analysis of cDNA for Bcl-2 and a hybrid bcl-2/immunoglobulin transcript resulting from the t(14;18) translocation. Cell 1986, 47:19–28
Chen-Levy Z, Nourse J, Cleary ML. The bcl-2 candidate proto-oncogene product is a 24-kilodalton integral-membrane protein highly expressed in lymphoid cell lines and lymphomas carrying the t(14:18) translocation. Mol Cell Biol 1989, 9:701–710
Hockenbery DM, Zutter M, Hickey W, Nahm M, Korsmeyer S. Bcl-2 protein is topographically restricted in tissues characterized by apoptotic cell death. Proc Natl Ac Sci USA 1991, 16:8479–8483
Lu QL, Elia G, Lucas S, Thomas JA. Bcl-2 proto-oncogene expression in Ebstein-Barr virus associated nasopharyngeal carcinoma. Cancer 1993, 53:29–35
Nunez G, London L, Hockenbery D, Alexander M, McKearn JP, Korsmeyer SJ. Deregulated Bcl-2 gene-expression selectively prolongs survival of growth factor-deprived hemopoietic cell lines. J Immumol 1991, 144:3602–3610
Bissonette RP, Echeverri F, Mahboubi A, Green DR. Apoptotic cell death induced by c-myc is inhibited by bcl-2. Nature 1992, 359:552–554
Fanidi A, Harrington EA, Evan GI. Cooperative interaction between c-myc and bcl-2 proto-oncogenes. Nature 1992, 359:554–556
Hickman JA. Apoptosis induced by anti-cancer drugs. Cancer Metast Rev 1992, 11:121–139
Dive C, Hickman JA. Drug-target interactions: Only first step in the commitment to programmed cell death?. Br J Cancer 1991, 64:192–196
O’Connor PM, Wassermann K, Sarang M, Magrath I, Bohr VA, Kohn KW. Relation between DNA crosslinks, cell-cycle, and apoptosis in Burkitt’s lymphoma cell lines differing in sensitivity to nitrogen mustard. Cancer Res 1991, 51:6550–6557
Lowe SW, Ruley HE, Jacks T, Housman DE. p53-dependent apoptosis modulates the cytotoxicity of anti-cancer agents. Cell 1993, 74:957–967
Miyashita T, Reed JC. Bcl-2 oncoprotein blocks chemotherapy-induced apoptosis in a human leukemia cell line. Blood 1990, 81:151–157
Sentman CL, Shutter JR, Hockenbery D, Kanagawa O, Korsmeyer SJ. Bcl-2 inhibits multiple forms of apoptosis but not negative selection in thymocytes. Cell 1991, 67:879–888
Smets LA, Van Den Berg J, Acton D, Top B, Van Rooij H, Verweij-Janssen. Bcl-2 expression and mitochondrial activity in leukemic cells with different sensitivity to glucocorticoid-induced apoptosis. Blood 1994, 84:1613–1619
Campos L, Rouault JP, Sabido O, Oriol P, Roubi N, Vasselon C, Archimbaud E, Magaud JP, Guyotot D. High expression of bcl-2 protein in acute myeloid leukemia cells is associated with poor respons to chemotherapy. Blood 1993, 81:3091–3096
Porwit-MacDonald A, Ivory K, Wilkinson S, Wheatley K, Wong L, Janossy G. Bcl-2 protein expression in normal human bone marrow precursors and in acute myelogenous leukemia. Leukemia 1995, 9:1191–1198
Reed JC. Bcl2: prevention of apoptosis as a mechanism of drug resistance. Hematol Oncol Clin North Am 1995, 9:451–473
Lotem J, Sachs L. Control of sensitivity to induction of apoptosis in myeloid leukemic cells by differentation and bcl-2 dependent and independent pathways. Cell Growth Differ 1994, 5:321–327
Gala JL, Vermeylen C, Cornu G, Ferrant A, Michaux JL, Philippe M, Martiat P. High expression of bcl-2 is the rule in acute lymphoblastic leukemia, except in Burkitt subtype at presentation, and is not correlated with the prognosis. Ann Hematol 1994, 69:17–24
Maung ZT, Maclean ER, Reid MM, Pearson AD, Proctor SJ, Hamilton PJ, Hall AG. The relationship between bcl-2 expression and response to chemotherapy in acute leukaemia. Br J Haematol 1994, 88:105–109
Coustan-Smith E, Kitanaka A, Pui CH, McNinch L, Evans WE, Raimondi SC, Behm FG, Arico M, Campana D. Clinical relevance of bcl-2 over-expression in childhood acute lymphoblastic leukemia. Blood 1996, 87:1140–1146
Kaspers GJL, Veerman AJP, Pieters R, Broekema GJ, Huismans DR, Kazemier KM, Loonen AH, Rottier MMA, Van Zantwijk CH, Hahlen K, Van Wering ER. Mononuclear cells contaminating leukemic samples tested for cellular drug resistance using the methyl-thiazol-tetrazolium assay. Br J Cancer 1994, 10:1047–1052
Castle VP, Heidelberger KP, Bromberg J, Ou X, Dole M, Nunez G. Expression of the apoptosis-surpressing gen bcl-2 in neuroblastoma is associated with unfavorable histology and N-myc amplification. Am J Path 1993, 143:1543–1550
McDonnell TJ, Troncoso P, Brisbay SM, Logothetis C, Ching LWK, Hsieh JT, Tu SM, Campbell ML. Expression of the protooncogene bcl-2 in the prostate and its association with emergence of androgen-independent prostate cancer. Cancer Res 1992, 52:6940–6944
Reed JC, Kitada S, Takayama S, Miyashita T. Regulation of chemoresistance by the bcl-2 oncoprotein in non-Hodgkin’s lymphoma and lymphoblastic leukemia cell lines. Ann Oncol 1994, 5:61–65
Klumper E, Pieters R, Veerman AJP, Huismans DR, Loonen AH, Hählen K, Kaspers GJL, Van Wering ER, Hartmann R, Henze G. In vitro cellular drug resistance in children with relapsed/refractory acute lymphoblastic leukemia. Blood 1995, 86:3681–3688
Salomons GS, Brady HJM, Verwijs-Janssen M, Van Den Berg JD, Hart AAM, Van Den Berg H, Behrendt H, Hählen K, Smets L. The Baxα: bcl-2 ratio modulates the respons to dexamethasone in leukemic cells and is highly variable in childhood acute leukemia. Fnt J Cancer 1997, 71:959–965
Findley HW, Gu I, Yeager AM, Zhou M. Expression and regulation of bcl-2, bcl-xl and bax correlate with p53 status and sensitivity to apoptosis in childhood acute lymphoblastic leukemia. Blood 1997, 89:2986–2993
Kaspers GJL, Pieters R, Klumper E, De Waal FC, Veerman AJP. Glucocorticoid resistance in childhood leukemia. Leuk & Lymphoma 1994, 13:187–201
Schena M, Larsson LG, Gottardi D, Gaidano G, Carlsson M, Nilsson K, Caligaris-Cappio F. Growth and differentiation-associated expression of bcl-2 in B-chronic lymphocytic leukemia cells. Blood 1992, 11:2981–2989
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1999 Springer Science+Business Media New York
About this chapter
Cite this chapter
Haarman, E.G. et al. (1999). BCL-2 Expression in Childhood Leukemia Versus Spontaneous Apoptosis, Drug Induced Apoptosis, and in vitro Drug Resistance. In: Kaspers, G.J.L., Pieters, R., Veerman, A.J.P. (eds) Drug Resistance in Leukemia and Lymphoma III. Advances in Experimental Medicine and Biology, vol 457. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-4811-9_35
Download citation
DOI: https://doi.org/10.1007/978-1-4615-4811-9_35
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4613-7180-9
Online ISBN: 978-1-4615-4811-9
eBook Packages: Springer Book Archive