Annals of Hematology

, Volume 84, Supplement 1, pp 47–53 | Cite as

Decitabine activates specific caspases downstream of p73 in myeloid leukemia

Original Article

Abstract

The demethylating effect of 5-aza-2′ deoxycytidine (decitabine, DAC) has been well characterized. The molecular events downstream of methylation inhibition are less well known. Here, DAC was shown to induce apoptosis in acute myeloid leukemia (AML) cells (p53 mutant and wild type) but not in epithelial or normal peripheral blood mononuclear cells. Apoptosis was characterized by activation of the mitochondrial but not the receptor death pathway, as demonstrated by the release of cytochrome c and loss of mitochondrial membrane potential. Western blotting and enzyme assays showed that caspase-3, but not caspase-6 or caspase-8, were activated. Decitabine induced expression of the cell cycle inhibitor p21, arresting AML cell lines in G1 of the cell cycle. Expression of p21 was induced irrespective of the methylation status of its promoter, mediated instead via reexpression of the tumor suppressor p73, an upstream regulator of p21. The promoter of p73 was hypermethylated in AML cell lines in vitro and in primary AML cells ex vivo but not in DAC-resistant epithelial cells. In conclusion, DAC acts on leukemic myeloid cells via caspase activation, which may be dependent on demethylation of the hypermethylated p73 promoter and consequent reexpression of p73.

Keywords

Acute myeloid leukemia (AML) Apoptosis Hypermethylation Caspases p73 p21 

References

  1. 1.
    Aparicio A, Eads CA, Leong LA, Laird PW, Newman EM, Synold TW, Baker SD, Zhao M, Weber JS (2003) Phase I trial of continuous infusion 5-aza-2′-deoxycytidine. Cancer Chemother Pharmacol 51:231–239PubMedGoogle Scholar
  2. 2.
    Chylicki K, Ehinger M, Svedberg H, Gullberg U (2000) Characterization of the molecular mechanisms for p53-mediated differentiation. Cell Growth Differ 11:561–571PubMedGoogle Scholar
  3. 3.
    Daskalakis M, Nguyen TT, Nguyen C, Guldberg P, Köhler G, Wijermans P, Jones PA, Lübbert M (2002) Demethylation of a hypermethylated p16/INK4B gene in patients with myeloblastic syndrome by 5-Aza-2′-deoxycytidine (decitabine) treatment. Blood 100:2957–2964CrossRefPubMedGoogle Scholar
  4. 4.
    Dotto GP (2000) p21(WAF1/Cip1): more than a break to the cell cycle? Biochim Biophys Acta 1471:43–56Google Scholar
  5. 5.
    el-Deiry WS, Tokino T, Velculescu VE, Levy DB, Parsons R, Trent JM, Lin D, Mercer WE, Kinzler KW, Vogelstein B (1993) WAF1, a potential mediator of p53 tumour suppression. Cell 75:817–825CrossRefPubMedGoogle Scholar
  6. 6.
    Fleckenstein DS, Uphoff CC, Drexler HG, Quentmeier H (2002) Detection of p53 gene mutations by single strand conformational polymorphism (SSCP) in human acute myeloid leukemia-derived cell lines. Leuk Res 26:207–214CrossRefPubMedGoogle Scholar
  7. 7.
    Fournel M, Sapieha P, Beaulieu N, Besterman JM, MacLeod AR (1999) Down-regulation of human DNA-(cytosine-5) methyltransferase induces cell cycle regulators p16(ink4A) and p21(WAF/Cip1) by distinct mechanisms. J Biol Chem 274:24250–24256CrossRefPubMedGoogle Scholar
  8. 8.
    Green DR, Reed JC (1998) Mitochondria and apoptosis. Science 281:1309–1312CrossRefPubMedGoogle Scholar
  9. 9.
    Irwin MS, Kondo K, Marin MC, Cheng LS, Hahn WC, Kaelin WG Jr (2003) Chemosensitivity linked to p73 function. Cancer Cell 3:403–410CrossRefPubMedGoogle Scholar
  10. 10.
    Issa JP, Garcia-Manero G, Giles FJ, Mannari R, Thomas D, Faderl S, Bayar E, Lyons J, Rosenfeld CS, Cortes J, Kantarjian HM (2003) Phase I study of low-dose prolonged exposure schedules of the hypomethylating agent 5-aza-2′-deoxycytidine (Decitabine) in hematopoietic malignancies. Blood 6:6Google Scholar
  11. 11.
    Jones PA, Baylin SB (2002) The fundamental role of epigenetic events in cancer. Nat Rev Genet 3:415–428CrossRefPubMedGoogle Scholar
  12. 12.
    Kantarjian HM, O'Brien S, Cortes J, Giles FJ, Faderl S, Issa JP, Garcia-Manero G, Rios MB, Shan J, Andreeff M, Keating M, Talpaz M (2003) Results of decitabine (5-aza-2′deoxycytidine) therapy in 130 patients with chronic myelogenous leukemia. Cancer 98:522–528CrossRefPubMedGoogle Scholar
  13. 13.
    Kawano S, Miller CW, Gombart AF, Bartram CR, Matsuo Y, Asou H, Sakashita A, Said J, Tatsumi E, Koeffler HP (1999) Loss of p73 gene expression in leukemias/lymphomas due to hypermethylation. Blood 94:1113–1120PubMedGoogle Scholar
  14. 14.
    Lavelle D, Chen YH, Hankewych M, DeSimone J (2001) Histone deacetylase inhibitors increase p21(WAF1) and induce apoptosis of human myeloma cell lines independent of decreased IL-6 receptor expression. Am J Hematol 68:170–178CrossRefPubMedGoogle Scholar
  15. 15.
    Melino G, Bernassola F, Ranalli M, Yee K, Zong WX, Corazzari M, Knight RA, Green DR, Thompson C, Vousden KH (2004) p73 induces apoptosis via PUMA transactivation and Bax mitochondrial translocation. J Biol Chem 279:8076–8083CrossRefPubMedGoogle Scholar
  16. 16.
    Milutinovic S, Knox JD, Szyf M (2000) DNA methyltransferase inhibition induces the transcription of the tumour suppressor p21(WAF1/CIP1/sdi1). J Biol Chem 275:6353–6359CrossRefPubMedGoogle Scholar
  17. 17.
    Robertson KD (2001) DNA methylation, methyltransferases, and cancer. Oncogene 20:3139–3155CrossRefPubMedGoogle Scholar
  18. 18.
    Roman-Gomez J, Castillejo J-A, Jimenez A, Gonzalez MG, Moreno F, Rodriguez MdC, Barrios M, Maldonado J, Torres A (2002) 5′ CpG island hypermethylation is associated with transcriptional silencing of the p21CIP/WAF1/SDI1 gene and confers poor prognosis in acute lymphoblastic leukemia. Blood 99:2291–2296CrossRefPubMedGoogle Scholar
  19. 19.
    Schmelz K, Wagner M, Dörken B, Tamm I (2005) 5-Aza-2′-deoxycytidine induces p21WAF expression by demethylation of p73 leading to p53-independent apoptosis in myeloid leukaemia. Int J Cancer 114:683–695CrossRefPubMedGoogle Scholar
  20. 20.
    Stirewalt DL, Clurman B, Appelbaum FR, Willman CL, Radich JP et al (1999) p73 mutations and expression in adult de novo acute myelogenous leukemia. Leukemia 13:985–990CrossRefPubMedGoogle Scholar
  21. 21.
    Terui T, Murakami K, Takimoto R, Takahashi M, Takada K, Murakami T, Minami S, Matsunaga T, Takayama T, Kato J, Niitsu Y (2003) Induction of PIG3 and NOXA through acetylation of p53 at 320 and 373 lysine residues as a mechanism for apoptotic cell death by histone deacetylase inhibitors. Cancer Res 63:8948–8954PubMedGoogle Scholar
  22. 22.
    Wijermans P, Lubbert M, Verhoef G, Bosly A, Ravoet C, Andre M, Ferrant A (2000) Low-dose 5-aza-2′deoxycytidine, a DNA hypomethylating agent, for the treatment of high-risk myelodysplastic syndrome: a multicenter phase II study in elderly patients. J Clin Oncol 18:956–962PubMedGoogle Scholar
  23. 23.
    Yang A, Kaghad M, Caput D, McKeon F (2002) On the shoulders of giants: p63, p73 and the rise of p53. Trends Genet 18:90–95CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  1. 1.Department of Hematology and OncologyUniversitätsmedizin Berlin Charité, Campus VirchowBerlinGermany

Personalised recommendations