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Fludarabine-induced apoptosis of B chronic lymphocytic leukemia cells includes early cleavage of p27kip1 by caspases

Leukemia volume 17, pages 1104–1111 (2003)Cite this article

Abstract

B-cell chronic lymphocytic leukemia (B-CLL) is characterized by the accumulation of growth arrested clonal B lymphocytes that undergo apoptosis when treated with fludarabine. To further explore the mechanism for the cell cycle arrest, we examined the expression and activity of cyclin-dependent kinases and inhibitors in primary B-CLL cells. We observed high levels of p27kip1, cyclin D2, cyclin E, cdk2, and cdk4 expression in freshly isolated B-CLL cells. Despite high levels of cyclins and cdks, little cdk2 or cdk4 activity was observed with p27kip1 in complex with cyclinD2/cdk4 and cyclin E/cdk2. Remarkably, when B-CLL cells were treated in vitro with fludarabine, p27kip1 underwent caspase-specific degradation accompanied by an increase in cdk4 activity. We conclude that the G0/G1 arrest of B-CLL cells may protect against apoptosis and that the decrease in p27kip1 expression by caspase cleavage may be a key step in chemotherapy-induced apoptosis in B-CLL.

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References

  1. Caligaris-Cappio F, Hamblin TJ . B-cell chronic lymphocytic leukemia: a bird of a different feather. J Clin Oncol 1999; 17: 399–408.

    Article  CAS  PubMed  Google Scholar 

  2. Klein A, Miera O, Bauer O, Golfier S, Schriever F . Chemosensitivity of B cell chronic lymphocytic leukemia and correlated expression of proteins regulating apoptosis, cell cycle and DNA repair. Leukemia 2000; 14: 40–46.

    Article  CAS  PubMed  Google Scholar 

  3. Robertson LE, Plunkett W, McConnell K, Keating MJ, McDonnell TJ . Bcl-2 expression in chronic lymphocytic leukemia and its correlation with the induction of apoptosis and clinical outcome. Leukemia 1996; 10: 456–459.

    CAS  PubMed  Google Scholar 

  4. Stoetzer OJ, Pogrebniak A, Scholz M, Pelka-Fleischer R, Gullis E, Darsow M et al. Drug-induced apoptosis in chronic lymphocytic leukemia. Leukemia 1999; 13: 1873–1880.

    Article  CAS  PubMed  Google Scholar 

  5. Vrhovac R, Delmer A, Tang RP et al. Prognostic significance of the cell cycle inhibitor p27kip1 in chronic B-cell lymphocytic leukemia. Blood 1998; 91: 4694–4700.

    CAS  PubMed  Google Scholar 

  6. Polyak K, Kato JY, Solomon M, Sherr CJ, Massague J, Roberts JM, Koff A . p27Kip1, a cyclin-Cdk inhibitor, links transforming growth factor-β and contact inhibition to cell cycle arrest. Genes and Dev 1994; 8: 9–22.

    Article  CAS  PubMed  Google Scholar 

  7. Sgambato A, Cittadini A, Faraglia B, Weinstein IB . Multiple functions of p27(Kip1) and its alterations in tumor cells: a review. J Cell Physiol 2000; 183: 18–27.

    Article  CAS  PubMed  Google Scholar 

  8. Bartek J, Bartkova J, Lukas J . The retinoblastoma protein pathway in cell cycle control and cancer. Exp Cell Res 1997; 237: 1–6.

    Article  CAS  PubMed  Google Scholar 

  9. Bouchard C, Thieke K, Maier A, Saffrich R, Hanley-Hyde J, Ansorge W et al. Direct induction of cyclin D2 by Myc contributes to cell cycle progression and sequestration of p27. EMBO J 1999; 18: 5321–5333.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Rivard N, L'Allemain G, Bartek J, Pouyssegur J . Abrogation of p27Kip1 by cDNA antisense suppresses quiescence (G0 state) in fibroblasts. J Biol Chem 1996; 271: 18337–18341.

    Article  CAS  PubMed  Google Scholar 

  11. Craig C, Wersto R, Kim M, Ohri E, Li Z, Katayose D et al. A recombinant adenovirus expressing p27Kip1 induces cell cycle arrest and loss of cyclin-Cdk activity in human breast cancer cells. Oncogene 1997; 14: 2283–2289.

    Article  CAS  PubMed  Google Scholar 

  12. Fero ML, Randel E, Gurley KE, Roberts JM, Kemp CJ . The murine gene p27Kip1 is haplo-insufficient for tumour suppression. Nature 1998; 396: 177–180.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Steeg PS, Abrams JS . Cancer prognostics: past, present, and p27. Nat Med 1997; 3: 152–154.

    Article  CAS  PubMed  Google Scholar 

  14. Sanchez-Beato M, Saez AI, Martinez-Montero JC et al. Cyclin-dependent kinase inhibitor p27KIP1 in lymphoid tissue: p27KIP1 expression is inversely proportional to the proliferative index. Am J Pathol 1997; 151: 151–160.

    CAS  PubMed  PubMed Central  Google Scholar 

  15. Hiromura K, Pippin JW, Fero ML, Roberts JM, Shankland SJ . Modulation of apoptosis by the cyclin-dependent kinase inhibitor p27(Kip1). J Clin Invest 1999; 103: 597–604.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Chandra J, Gilbreath J, Freireich EJ et al. Protease activation is required for glucocorticoid-induced apoptosis in chronic lymphocytic leukemic lymphocytes. Blood 1997; 90: 3673–3681.

    CAS  PubMed  Google Scholar 

  17. Pettitt AR, Cawley JC . Caspases influence the mode but not the extent of cell death induced by purine analogues in chronic lymphocytic leukaemia. Br J Haematol 2000; 109: 800–804.

    Article  CAS  PubMed  Google Scholar 

  18. Levkau B, Koyama H, Raines EW, Clurman BE, Herren B, Orth K et al. Cleavage of p21Cip1/Waf1 and p27Kip1 mediates apoptosis in endothelial cells through activation of Cdk2: role of a caspase cascade. Mol Cell 1998; 1: 553–563.

    Article  CAS  PubMed  Google Scholar 

  19. Boutillier AL, Trinh E, Loeffler JP . Caspase-dependent cleavage of the retinoblastoma protein is an early step in neuronal apoptosis. Oncogene 2000; 19: 2171–2178.

    Article  CAS  PubMed  Google Scholar 

  20. Loubat A, Rochet N, Turchi L, Rezzonico R, Far DF, Auberger P et al. Evidence for a p23 caspase-cleaved form of p27[KIP1] involved in G1 growth arrest. Oncogene 1999; 18: 3324–3333.

    Article  CAS  PubMed  Google Scholar 

  21. Delmer A, Ajchenbaum-Cymbalista F, Tang R, Ramond S, Faussat AM, Marie JP, Zittoun R . Over-expression of cyclin D1 in chronic B-cell malignancies with abnormality of chromosome 11q13. Br J Haematol 1995; 89: 798–804.

    Article  CAS  PubMed  Google Scholar 

  22. Bellosillo B, Dalmau M, Colomer D, Gil J . Involvement of CED-3/ICE proteases in the apoptosis of B-chronic lymphocytic leukemia cells. Blood 1997; 89: 3378–3384.

    CAS  PubMed  Google Scholar 

  23. Polyak K, Lee MH, Erdjument-Bromage H, Koff A, Roberts JM, Tempst P, Massague J . Cloning of p27Kip1, a cyclin-dependent kinase inhibitor and a potential mediator of extracellular antimitogenic signals. Cell 1994; 78: 59–66.

    Article  CAS  PubMed  Google Scholar 

  24. Tan X, Wang JY . The caspase-RB connection in cell death. Trends Cell Biol 1998; 8: 116–120.

    Article  CAS  PubMed  Google Scholar 

  25. Canhoto AJ, Chestukhin A, Litovchick L, DeCaprio JA . Phosphorylation of the retinoblastoma-related protein p130 in growth-arrested cells. Oncogene 2000; 19: 5116–5122.

    Article  CAS  PubMed  Google Scholar 

  26. Nakayama K, Ishida N, Shirane M, Inomata A, Inoue T, Shishido N et al. Mice lacking p27(Kip1) display increased body size, multiple organ hyperplasia, retinal dysplasia, and pituitary tumors. Cell 1996; 85: 707–720.

    Article  CAS  PubMed  Google Scholar 

  27. Coats S, Flanagan WM, Nourse J, Roberts JM . Requirement of p27Kip1 for restriction point control of the fibroblast cell cycle. Science 1996; 272: 877–880.

    Article  CAS  PubMed  Google Scholar 

  28. Ferguson KL, Callaghan SM, O'Hare MJ, Park DS, Slack RS . The Rb-CDK4/6 signaling pathway is critical in neural precursor cell cycle regulation [In Process Citation]. J Biol Chem 2000; 275: 33593–33600.

    Article  CAS  PubMed  Google Scholar 

  29. Zhang X, Wharton W, Donovan M, Coppola D, Croxton R, Cress WD, Pledger WJ . Density-dependent growth inhibition of fibroblasts ectopically expressing p27(kip1). Mol Biol Cell 2000; 11: 2117–2130.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Wang X, Gorospe M, Huang Y, Holbrook NJ . p27Kip1 overexpression causes apoptotic death of mammalian cells. Oncogene 1997; 15: 2991–2997.

    Article  CAS  PubMed  Google Scholar 

  31. Katayose Y, Kim M, Rakkar AN, Li Z, Cowan KH, Seth P . Promoting apoptosis: a novel activity associated with the cyclin- dependent kinase inhibitor p27. Cancer Res 1997; 57: 5441–5445.

    CAS  PubMed  Google Scholar 

  32. Dijkers PF, Medema RH, Pals C, Banerji L, Thomas NS, Lam EW et al. Forkhead transcription factor FKHR-L1 modulates cytokine-dependent transcriptional regulation of p27(KIP1). Mol Cell Biol 2000; 20: 9138–9148.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Dimanche-Boitrel MT, Micheau O, Hammann A, Haugg M, Eymin B, Chauffert B, Solary E . Contribution of the cyclin-dependent kinase inhibitor p27KIP1 to the confluence-dependent resistance of HT29 human colon carcinoma cells. Int J Cancer 1998; 77: 796–802.

    Article  CAS  PubMed  Google Scholar 

  34. Masuda A, Osada H, Yatabe Y, Kozaki K, Tatematsu Y, Takahashi T, Hida T . Protective function of p27(KIP1) against apoptosis in small cell lung cancer cells in unfavorable microenvironments. Am J Pathol 2001; 158: 87–96.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Schoenfeld AR, Parris T, Eisenberger A, Davidowitz EJ, De Leon M, Talasazan F et al. The von Hippel-Lindau tumor suppressor gene protects cells from UV-mediated apoptosis. Oncogene 2000; 19: 5851–5857.

    Article  CAS  PubMed  Google Scholar 

  36. Eymin B, Sordet O, Droin N, Munsch B, Haugg M, Van de Craen M et al. Caspase-induced proteolysis of the cyclin-dependent kinase inhibitor p27Kip1 mediates its anti-apoptotic activity. Oncogene 1999; 18: 4839–4847.

    Article  CAS  PubMed  Google Scholar 

  37. Park DS, Morris EJ, Padmanabhan J, Shelanski ML, Geller HM, Greene LA . Cyclin-dependent kinases participate in death of neurons evoked by DNA-damaging agents. J Cell Biol 1998; 143: 457–467.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Lundberg AS, Weinberg RA . Functional inactivation of the retinoblastoma protein requires sequential modification by at least two distinct cyclin-cdk complexes. Mol Cell Biol 1998; 18: 753–761.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Johnson DG, Schneider-Broussard R . Role of E2F in cell cycle control and cancer. Front Biosci 1998; 27: 447–448.

    Article  Google Scholar 

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Acknowledgements

We thank Nathalie Rochet and Gilles Ponzio for helpful discussions. This work was supported in part by Ligue contre le Cancer, Comité de Paris.

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Authors and Affiliations

  1. Hematology Department, INSERM 9912-EA1517, Hotel-Dieu, APHP, Paris, France

    L Sanhes, R Tang, A Delmer & F Ajchenbaum-Cymbalista

  2. Dana Farber Cancer Institute, Boston, MA, USA

    J A DeCaprio

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  1. L Sanhes
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Sanhes, L., Tang, R., Delmer, A. et al. Fludarabine-induced apoptosis of B chronic lymphocytic leukemia cells includes early cleavage of p27kip1 by caspases. Leukemia 17, 1104–1111 (2003). https://doi.org/10.1038/sj.leu.2402895

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