Role of BCL-2 and cell cycle regulatory proteins for corticosensitivity assessment in childhood acute lymphoblastic leukaemia
N. Baghdassarian
Laboratoire de Cytologie Analytique, Université Claude Bernard, MESRT JE 1879,
Search for more papers by this authorY. Bertrand
Laboratoire de Cytologie Analytique, Université Claude Bernard, MESRT JE 1879,
Service d'Hématologie Pédiatrique, Hôpital Debrousse,
Search for more papers by this authorP. Ffrench
Laboratoire central d'Hématologie et de Cytogénétique, Hôpital E. Herriot, and
Search for more papers by this authorP. A. Bryon
Laboratoire de Cytologie Analytique, Université Claude Bernard, MESRT JE 1879,
Laboratoire central d'Hématologie et de Cytogénétique, Hôpital E. Herriot, and
Search for more papers by this authorM. Ffrench
Laboratoire de Cytologie Analytique, Université Claude Bernard, MESRT JE 1879,
Laboratoire central d'Hématologie et de Cytogénétique, Hôpital E. Herriot, and
Search for more papers by this authorN. Baghdassarian
Laboratoire de Cytologie Analytique, Université Claude Bernard, MESRT JE 1879,
Search for more papers by this authorY. Bertrand
Laboratoire de Cytologie Analytique, Université Claude Bernard, MESRT JE 1879,
Service d'Hématologie Pédiatrique, Hôpital Debrousse,
Search for more papers by this authorP. Ffrench
Laboratoire central d'Hématologie et de Cytogénétique, Hôpital E. Herriot, and
Search for more papers by this authorP. A. Bryon
Laboratoire de Cytologie Analytique, Université Claude Bernard, MESRT JE 1879,
Laboratoire central d'Hématologie et de Cytogénétique, Hôpital E. Herriot, and
Search for more papers by this authorM. Ffrench
Laboratoire de Cytologie Analytique, Université Claude Bernard, MESRT JE 1879,
Laboratoire central d'Hématologie et de Cytogénétique, Hôpital E. Herriot, and
Search for more papers by this authorAbstract
Results of treatment in childhood acute lymphoblastic leukaemia (ALL) remain unsatisfactory because relapses occur even after high-dose chemotherapy. Corticosensitivity is used in numerous therapeutic trials as a prognostic factor for treatment choice. The aim of this study was to evaluate the role of cell cycle regulatory protein expression before and during the first 48 h of corticotherapy for predicting corticosensitivity. Fifty-two children presenting with ALL were studied at diagnosis and during the first 48 h of treatment for cell proliferation and apoptosis level by measurement of DNA content, and for expression of several cell proliferation regulatory proteins by means of Western blot. Glucocorticoids induced a significant decrease in the percentage of cells in S-phase and in CDK1, CDK4 and CDK6 expression and an increase in the percentage of cells in subG1 peak. Two criteria for corticosensitivity were used: (i) the number of blast cells after 7 d of treatment with a threshold at 1 × 109/l (usual criterion), (ii) the J8/J1 blast cell ratio, which is independent from initial leucocytosis. Bcl-2 expression at diagnosis was the best predictive variable for the usual corticosensitivity criterion in B- and T-cell ALL. For the second criterion, in B-cell ALL, p21waf1 expression at diagnosis was the sole (albeit poorly) predictive variable, whereas bcl-2 remained of high interest in T-cell ALL. Interestingly, these proteins, bcl-2 and p21waf1, are associated with prolonged cell lifespan and their increased expression is often linked to poor response to cytotoxic drugs. Such preliminary results call for subsequent studies on large independent sets of T-cell and B-cell lineage ALL in order to confirm the J8/J1 blast cell ratio value as well as the role of bcl-2 and p21waf1 expression in predicting corticosensitivity.
References
- 1 Ajchenbaum, F., Ando, K., De Caprio, J.A. & Griffin, J.D. (1993) Independent regulation of human d-type cyclin gene expression during G1 phase in primary human T lymphocytes. Journal of Biological Chemistry, 268, 4113 – 4119.
- 2 Alnemri, E.S., Fernandes, T.F., Haldar, S., Croce, C.M. & Litwack, G. (1992) Involvement of BCL-2 in glucocorticoid-induced apoptosis of human pre-B-leukemias. Cancer Research, 52, 491 – 495.
- 3 Baghdassarian, N., Catallo, R., Mahly, M.A., Ffrench, P., Chizat, F., Bryon, P.A. & Ffrench, M. (1998) Glucocorticoids induce G1 as well as S-phase lengthening in normal human stimulated lymphocytes: differential effects on cell-cycle regulatory proteins. Experimental Cell Research, 240, 263 – 273.
- 4 Baghdassarian, N., Peiretti, A., Devaux, E., Bryon, P.A. & Ffrench, M. (1999) Involvement of p27kip1 in the G1 and S-phase lengthening mediated by glucocorticoids in normal human lymphocytes. Cell Growth and Differentiation, 10, 405 – 412.
- 5 Beato, M. (1989) Gene regulation by steroid hormones. Cell, 56, 335 – 344.
- 6 Bennett, J.M., Catovsky, D., Daniel, M.T., Flandrin, G., Galton, D.A.G., Gralnick, H.R. & Sultan, C. (1981) The French–American–British (FAB) co-operative group. The morphological classification of acute lymphoblastic leukaemia: concordance among observers and clinical correlations. British Journal of Haematology, 47, 553 – 561.
- 7 Boumpas, D.T., Anastassiou, E.D., Older, S.A., Tsokos, G.C., Nelson, D.L. & Balow, J.E. (1991) Dexamethasone inhibits human interleukin 2 but not interleukin 2 receptor gene expression in vitro at the level of nuclear transcription. Journal of Clinical Investigation, 87, 1739 – 1747.
- 8 Campana, D., Coustan-Smith, E., Manabe, A., Buschle, M., Raimondi, S.C., Behm, F.G., Ashmun, R., Arico, M., Biondi, A. & Pui, C.H. (1993) Prolonged survival of B-lineage acute lymphoblastic leukemia cells is accompanied by overexpression of bcl-2 protein. Blood, 81, 1025 – 1031.
- 9 Corroyer, S., Nabeyrat, E. & Clement, A. (1997) Involvement of the cell cycle inhibitor CIP1/WAF1 in lung alveolar epithelial cell growth arrest induced by glucocorticoids. Endocrinology, 138, 3677 – 3685.
- 10 Darzynkiewicz, Z., Bruno, S., Del Biro, G., Gorezyca, W., Motz, M.A., Lassota, P. & Traganos, F. (1992) Features of apoptotic cells measured by flow cytometry. Cytometry, 13, 795 – 808.
- 11 Draetta, G. & Beach, D. (1988) Activation of cdc2 protein kinase during mitosis in human cells: cell-cycle dependent phosphorylation and subunit rearrangement. Cell, 54, 17 – 26.
- 12 Eastman-Recks, S.B. & Vedeckis, W.V. (1986) Glucocorticoid inhibition of c-myc, c-myb, c-ki-ras in a mouse lymphoma cell line. Cancer Research, 46, 2457 – 2462.
- 13 Ernst, P. & Killmann, S.-A. (1970) Perturbation of generation cycle of human leukemic blast cells by cytostatic therapy in vivo: effect of corticosteroids. Blood, 36, 689 – 696.
- 14 Fan, S., Chang, J.K., Smith, M.L., Duba, D., Fornace, A.J. & O'Connor, P.M. (1997) Cells lacking Cip1/Waf1 genes exhibit preferential sensitivity to cisplatin and nitrogen mustard. Oncogene, 14, 2127 – 2136.
- 15 Fanger, B.O., Schreifer, J. & Cidlowski, A. (1987) Glucocorticoids increase the length of G2 and M phases of HeLaS3 cell cycle. Journal of Steroid Biochemistry, 28, 345 – 347.
- 16 Ffrench, M., Bryon, P.A., Fière, D., Van Vu, H., Gentilhomme, O., Adeleine, P. & Viala, J.J. (1985) Cell-cycle, protein content and nuclear size in acute myeloid leukemia. Cytometry, 6, 47 – 53.
- 17 Hara, E., Smith, R., Parry, D., Tahara, H., Stone, S. & Peters, G. (1996) Regulation of p16CDKN2 expression and its implication for cell immortalization and senescence. Molecular and Cellular Biology, 16, 859 – 867.
- 18 Kato, G.J., Quddus, F.F., Shuster, J.J., Boyett, J., Pullen, J.D., Borowitz, M.J., Whitehead, V.M., Crist, W.M. & Leventhal, B.G. (1993) High glucocorticoid receptor content of leukemic blasts is a favorable prognostic factor in childhood acute lymphoblastic leukemia. Blood, 82, 2304 – 2309.
- 19 Mastrangelo, R., Malandrino, R., Riccardi, R., Longo, P., Ranelletti, F.O. & Iacobelli, S. (1980) Clinical implications of glucocorticoid receptor studies in childhood acute lymphoblastic leukemia. Blood, 56, 1036 – 1040.
- 20 Matsushime, H., Ewen, M.E., Strom, D.K., Kato, J.Y., Hanks, S.K., Roussel, M.F. & Sherr, C.J. (1992) Identification and properties of an atypical catalytic subunit (p34PSK–J3/cdk4) for mammalian D type G1 cyclins. Cell, 71, 323 – 334.
- 21 Mekki, Y., Catallo, R., Bertrand, Y., Manel, A.M., Ffrench, P., Baghdassarian, N., Duhaut, P., Bryon, P.A. & Ffrench, M. (1999) Enhanced expression of p16ink4a is associated with a poor prognosis in childhood acute lymphoblastic leukemia. Leukemia, 13, 181 – 189.
- 22 Meyerson, M. & Harlow, E. (1994) Identification of G1 kinase activity for cdk6, a novel cyclin D partner. Molecular and Cellular Biology, 14, 2077 – 2086.
- 23 Miyashita, T. & Reed, J.C. (1993) BCL-2 oncoprotein blocks chemotherapy-induced apoptosis in a human leukemia cell line. Blood, 81, 151–.
- 24 Modiano, J.F., Domenico, J., Szepesi, A., Lucas, J.J. & Gelfand, E.W. (1994) Differential requirements for interleukin-2 distinguish the expression and activity of the cyclin-dependent kinases Cdk4 and Cdk2 in human T cells. Journal of Biological Chemistry, 269, 32972 – 32978.
- 25 Noda, A., Ning, Y., Venable, S.F., Pereira-Smith, O.M. & Smith, J.R. (1994) Cloning of senescent cell-derived inhibitors of DNA synthesis using an expression screen. Experimental Cell Research, 211, 90 – 98.
- 26 Nourse, J., Firpo, E., Flanagan, W.M., Coats, S., Polyak, K., Lee, M.-H., Massague, J., Crabtree, G.R. & Roberts, J.M. (1994) Interleukin-2-mediated elimination of the p27Kip1 cyclin-dependent kinase inhibitor prevented by rapamycin. Nature, 372, 570 – 573.
- 27 Oakley, R.H. & Cidlowski, J.A. (1993) Homologous down regulation of the glucocorticoid receptor: the molecular machinery. Critical Reviews in Eukaryote Gene Expression, 3, 63 – 68.
- 28 Pagano, M., Pepperkok, R., Lukas, J., Baldin, V., Ansorge, W., Bartek, J. & Draetta, G. (1993) Regulation of the cell cycle by the cdk2 protein kinase in cultured fibroblasts. Journal of Cell Biology, 121, 101 – 111.
- 29 Ramalingam, A., Hirai, A. & Thompson, E.A. (1997) Glucocorticoid inhibition of fibroblast proliferation and regulation of the cyclin kinase inhibitor p21waf1. Molecular Endocrinology, 11, 577 – 586.
- 30 Reisman, D. & Thompson, E.A. (1995) Glucocorticoid regulation of cyclin D3 gene transcription and mRNA stability in lymphoid cells. Molecular Endocrinology, 9, 1500 – 1509.
- 31 Rhee, K., Reisman, D., Bresnahan, W. & Thompson, E.A. (1995) Glucocorticoid regulation of G1 cyclin-dependent kinase genes in lymphoid cells. Cell Growth and Differentiation, 6, 691 – 698.
- 32 Salinovich, O. & Montelaro, R. (1986) Reversible staining and peptide mapping of protein transferred to nitrocellulose after separation by SDS-PAGE. Analytical Biochemistry, 156, 341 – 347.
- 33 Sherr, C.J. (1993) Mammalian G1 cyclins. Cell, 73, 1059 – 1065.
- 34 Sherr, C.J. & Roberts, J. (1995) Inhibitors of mammalian G1 cyclin-dependent kinases. Genes and Development, 9, 1149 – 1163.
- 35 Thyss, A., Suciu, S., Bertrand, Y., Mazingue, F., Robert, A., Vilmer, E., Mechinaud, F., Benoit, Y., Brock, P., Ferster, A., Lutz, P., Boutard, P., Marguerite, G., Plouvier, E., Michel, G., Plantaz, D., Munzer, M., Rialland, X., Chantraine, J.M., Norton, L., Solbu, G., Philippe, N. & Otten, J. (1997) Systemic effect of intrathecal methotrexate during the initial phase of treatment of childhood acute lymphoblastic leukemia. Journal of Clinical Oncology, 15, 1824 – 1830.
- 36 Uckun, F.M., Yang, Z., Sather, H., Steinherz, P., Nachman, J., Bostrom, B., Crotty, L., Sarquis, M., Ek, O., Zeren, T., Tubergen, D., Reaman, G. & Gaynon, P. (1997) Cellular expression of antiapoptotic bcl-2 oncoprotein in newly diagnosed childhood acute lymphoblastic leukemia: a children's cancer group study. Blood, 89, 3769 – 3777.
- 37 Wolowiec, D., Deviller, P., Simonin, D., Souchier, C., Rimokh, R., Benchaib, M., Bryon, P.A. & Ffrench, M. (1995) Cdk1 is a marker of proliferation in human lymphoid cells. International Journal of Cancer, 61, 381 – 388.
- 38 Wolowiec, D., Mekki, Y., Ffrench, P., Manel, A.M., Bertrand, Y., Rimokh, R., Philippe, N., Bryon, P.A. & Ffrench, M. (1996) Differential expression of cell proliferation regulatory proteins in B- and T-lineage acute lymphoblastic leukaemias. British Journal of Haematology, 95, 518 – 523.
- 39 Yuh, Y.S. & Thompson, E.B. (1989) Glucocorticoid effect on oncogene/growth gene expression in human T lymphoblastic leukemic cell line CCRF-CEM. Journal of Biological Chemistry, 264, 10904 – 10910.
Citing Literature
