DNA Methyltransferase Inhibitors

Paving the Way for Epigenetic Cancer Therapeutics
  • Gregory K. Reid
  • A. Robert MacLeod
Part of the Medical Intelligence Unit book series (MIUN)


Regional hypermethylation and global hypomethylation coexist in cancer cells. Under-standing the mechanisms responsible for global hypomethylation and regional hypermethylation in cancer is required for the proper design of therapeutic strategies targeting the DNA methylation machinery. This chapter discusses different models explaining this paradox. Global hypomethylation is proposed to be associated with activation by demethylation of metastasis-associated genes. Thus, anticancer therapy directed at DNA methyltransferase might have the untoward effect of promoting metastasis. Inhibition of demethylase activity on the other hand could potentially inhibit metastasis. It is therefore im-portant to identify and characterize the enzymes responsible for global hypomethylation in cancer.


Acute Myeloid Leukemia Tumor Suppressor Gene Chronic Myelogenous Leukemia Nucleoside Analogue Aberrant Methylation 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


  1. 1.
    Cohen P. Protein kinases—the major drug targets of the twenty-first century? Nat Rev Drug Discov 2002; 1(4):309–15.PubMedCrossRefGoogle Scholar
  2. 2.
    Keshet I, Lieman-Hurwitz J, Cedar H. DNA methylation affects the formation of active chromatin. Cell 1986; 44:535–43.PubMedCrossRefGoogle Scholar
  3. 3.
    Li E, Beard C, Jaenisch R. Role for DNA methylation in genomic imprinting. Nature 1993; 366:362–65.PubMedCrossRefGoogle Scholar
  4. 4.
    Shemer R et al. Dynamic methylation adjustment and counting as part of imprinting mechanisms. Proc Natl Acad Sci USA 1996; 93:6371–76.PubMedCrossRefGoogle Scholar
  5. 5.
    Selig S, Ariel M, Goitein R et al. Regulation of mouse satellite DNA replication time. EMBO J 1988; 7:419–26.PubMedGoogle Scholar
  6. 6.
    Yeivin A, Razin A. Gene methylation patterns and expression. EXS 1993; 64:523.PubMedGoogle Scholar
  7. 7.
    Robertson KD. DNA methylation and chromatin unraveling the tangled web. Oncogene 2002;21(35):5361–79.PubMedCrossRefGoogle Scholar
  8. 8.
    Cameron EE, Bachman KE, Myohanen S et al. Synergy of demethylation and histone deacetylase inhibition in the re-expression of genes silenced in cancer. Nat Genet 1999; 21:103–107.PubMedCrossRefGoogle Scholar
  9. 9.
    Lapeyre JN, Becker FF. 5-Methylcytosine content of nuclear DNA during chemical hepatocarcinogenesis and in carcinomas which result. Biochem Biophys Res Commun 1979; 87(3):698–705.PubMedCrossRefGoogle Scholar
  10. 10.
    Ghoshal AK, Farber E. The induction of liver cancer by dietary deficiency of choline and methionine without added carcinogens.Google Scholar
  11. 11.
    Lombardi B, Shinozuka H. Enhancement of 2-acetylaminofluorene liver carcinogenesis in rats fed a choline-devoid diet. Int J Cancer 1979; 23(4):565–70.PubMedCrossRefGoogle Scholar
  12. 12.
    Feinberg AP, Gehrke CW, Kuo KC et al. Reduced genomic 5-methylcytosine content in human colonic neoplasia. Cancer Res 1988; 48(5):1159–61.PubMedGoogle Scholar
  13. 13.
    Feinberg AP, Vogelstein B. Alterations in DNA methylation in human colon neoplasia. Semin Surg Oncol 1987; 3(3):149–51.PubMedCrossRefGoogle Scholar
  14. 14.
    Goelz SE, Vogelstein B, Hamilton SR et al. Hypomethylation of DNA from benign and malignant human colon neoplasms. Science 1985; 228(4696):187–90.PubMedCrossRefGoogle Scholar
  15. 15.
    Land H, Parada LF, Weinberg RA. Cellular oncogenes and multistep carcinogenesis. Science 1983;222(4625):771–8.PubMedCrossRefGoogle Scholar
  16. 16.
    Feinberg AP, Vogelstein B. Hypomethylation of ras oncogenes in primary human cancers. Biochem Biophys Res Commun 1983; 111(1):47–54.PubMedCrossRefGoogle Scholar
  17. 17.
    Feinberg AP, Vogelstein B. Hypomethylation distinguishes genes of some human cancers from their normal counterparts. Nature 1983; 301(5895):89–92.PubMedCrossRefGoogle Scholar
  18. 18.
    Christman JK, Price P, Pedrinan L et al. Correlation between hypomethylation of DNA and expression of globin genes in Friend erythroleukemia cells. Eur J Biochem 1977; 81(1):53–61.PubMedCrossRefGoogle Scholar
  19. 19.
    Constantinides PG, Jones PA et al. Functional striated muscle cells from non-myoblast precursors following 5-azacytidine treatment. Nature 1977; 267(5609):364–6.PubMedCrossRefGoogle Scholar
  20. 20.
    Jones PA, Taylor SM. Cellular differentiation, cytidine analogs and DNA methylation. Cell 1980;20(1):85–93.PubMedCrossRefGoogle Scholar
  21. 21.
    Taylor EM, McFarlane RJ, Price C. Mol Gen Genet 1996;253.Google Scholar
  22. 22.
    Lal D, Som S, Friedman S. Mutat Res 1988;193.Google Scholar
  23. 23.
    Tamame M, Antequera F, Villanueva JR et al. Mol Cell Biol 1983; 3:2287.PubMedGoogle Scholar
  24. 24.
    Esteller M. CpG island hypermethylation and tumor suppressor genes: a booming present, a brighter future. Oncogene 2002; 21(35):5427–40.PubMedCrossRefGoogle Scholar
  25. 25.
    Myohanen SK, Baylin SB, Herman JG. Hypermethylation can selectively silence individual p16ink4aink4A alleles in neoplasia. Cancer Res 1998; 58(4):591–3.PubMedGoogle Scholar
  26. 26.
    Adams RL, McKay EL, Craig LM et al. Mouse DNA methylase: methylation of native DNA. Biochim Biophys Acta 1979; 561:345–57.PubMedGoogle Scholar
  27. 27.
    Bestor T, Laudano A, Mattaliano R et al. Cloning and sequencing of a cDNA encoding DNA methyltransferase of mouse cells. The carboxyl-terminal domain of the mammalian enzymes is related to bacterial restriction methyltransferases. J Mol Biol 1988; 203(4):971–83.PubMedCrossRefGoogle Scholar
  28. 28.
    Yoder JA, Soman NS, Verdine GL et al. DNA (cytosine-5)-methyltransferases in mouse cells and tissues. Studies with a mechanism-based probe. J Mol Biol 1997; 270:385–95.PubMedCrossRefGoogle Scholar
  29. 29.
    Okano M, Xie S, Li E. Cloning and characterization of a family of novel mammalian DNA (cytosine-5) methyltransferases. Nat Genet 1998; 19:219–20.PubMedCrossRefGoogle Scholar
  30. 30.
    Li E, Bestor TH, Jaenisch R. Targeted mutation of the DNA methyltransferase gene results in embryonic lethality. Cell 1992; 69:915–26.PubMedCrossRefGoogle Scholar
  31. 31.
    Okano M, Bell DW, Haber DA et al. DNA methyltransferases Dnmt3a and Dnmt3b are essential for de novo methylation and mammalian development. Cell 1999; 99:247–57.PubMedCrossRefGoogle Scholar
  32. 32.
    Robertson KD, Uzvolgyi E, Liang G et al. The human DNA methyltransferase (DNMTs) 1, 3a and 3b: coordinate mRNA expression in normal tissues and overexpression in tumors. Nucleic Acids Res 1999; 27:2291–98.PubMedCrossRefGoogle Scholar
  33. 33.
    Hansen RS, Wijmenga C, Luo P et al. The DNMT3B DNA methyltransferase gene is mutated in the ICF immunodeficiency syndrome. Proc Natl Acad Sci USA 1999; 96:14412–7.PubMedCrossRefGoogle Scholar
  34. 34.
    Beaulieu N, Morin S, Chute IC et al. An essential role for DNA methyltransferase DNMT3B in cancer cell survival. J Biol Chem 2002; 277(31):28176–81PubMedCrossRefGoogle Scholar
  35. 35.
    Galmarini CM, Mackey JR, Dumontet C. Nucleoside analogues and nucleobases in cancer treatment. Lancet Oncol 2002; 3(7):415–24.PubMedCrossRefGoogle Scholar
  36. 36.
    Juttermann R, Li E, Jaenisch R. Toxicity of 5-aza-2′-deoxycytidine to mammalian cells is mediated primarily by covalent trapping of DNA methyltransferase rather than DNA demethylation. Proc Natl Acad Sci USA 1999; 91:1797–801.Google Scholar
  37. 37.
    Baylin S, Bestor TH. Altered methylation patterns in cancer cell genomes: cause or consequence? Cancer Cell 2002; 1(4):299–305.PubMedCrossRefGoogle Scholar
  38. 38.
    Rouleau J, MacLeod AR, Szyf M. Regulation of the DNA methyltransferase by the Ras-AP-1 signaling pathway. J Biol Chem 1995; 270(4):1595–601PubMedCrossRefGoogle Scholar
  39. 39.
    MacLeod RA, Rouleau J, Szyf M. Regulation of DNA methylation by the Ras signaling. J Biol Chem 1995; 270:11327–337.PubMedCrossRefGoogle Scholar
  40. 40.
    MacLeod RA, Szyf M. Expression of antisense to DNA methyltransferase mRNA induces DNA demethylation and inhibits tumorigenesis. J Biol Chem 1995; 270:8037–43.PubMedCrossRefGoogle Scholar
  41. 41.
    Ramchandani S, MacLeod AR, Pinard M et al. Inhibition of tumorigenesis by a cytosine-DNA, methyltransferase, antisense oligodeoxynucleotide. Proc Natl Acad Sci USA 1997; 94(2):684–9PubMedCrossRefGoogle Scholar
  42. 42.
    Laird PW, Jackson-Grusby L, Fazeli A et al. Suppression of intestinal neoplasia by DNA hypomethylation. Cell 1995; 81:197–205.PubMedCrossRefGoogle Scholar
  43. 43.
    Hussussian CJ et al. Germline p16ink4a mutations in familial melanoma. Nat Genet 1994; 8:15–21.PubMedCrossRefGoogle Scholar
  44. 44.
    Gonzalez-Zulueta M et al. Methylation of the 5′ CpG island of the p16ink4a/CDKN2 tumor suppressor gene in normal and transformed human tissues correlates with gene silencing. Cancer Res 1995; 55:4531–5.PubMedGoogle Scholar
  45. 45.
    Fournel M, Sapieha P, Beaulieu N et al. Down-regulation of human DNA-(cytosine-5) methyltransferase induces cell cycle regulators p16ink4a(ink4A) and p21(WAF/Cip1) by distinct mechanisms. J Bio Chem 1999; 274:24250–56.CrossRefGoogle Scholar
  46. 46.
    Rhee I, Jair KW, Yen RW et al. CpG methylation is maintained in human cancer cells lacking DNMT1. Nature 2000; 404(6781):1003–7.PubMedCrossRefGoogle Scholar
  47. 47.
    Rhee I et al. DNMT1 and DNMT3b cooperate to silence genes in human cancer cells. Nature 2002; 416(6880):552–6.PubMedCrossRefGoogle Scholar
  48. 48.
    Robert MF, Morin S, Beaulieu N et al. DNMT1 is required to maintain CpG methylation and aberrant gene silencing in human cancer cells. Nat Genet 2003; 33(1):61–5.PubMedCrossRefGoogle Scholar
  49. 49.
    Opalinska JB, Gewirtz AM. Nucleic-acid therapeutics: basic principles and recent applications. Nat Rev Drug Discov 2002; 1(7):503–14.PubMedCrossRefGoogle Scholar
  50. 50.
    Bender CM, Pao MM, Jones PA. Inhibition of DNA methylation by 5-aza-2′-deoxycytidine suppresses the growth of human tumor cell lines. Cancer Res 1998; 58(1):95–101.PubMedGoogle Scholar
  51. 51.
    Bakin AV, Curran T. Role of DNA 5-methylcytosine transferase in cell transformation by fos. Science 1999; 283:387–90.PubMedCrossRefGoogle Scholar
  52. 52.
    Di Croce L, Raker VA, Corsaro M et al. Methyltransferase recruitment and DNA hypermethylation of target promoters by an oncogenic transcription factor. Science 2002; 295:1079–82.PubMedCrossRefGoogle Scholar
  53. 53.
    Chuang LS, Ian HI, Koh TW et al. Human DNA-(cytosine-5) methyltransferase-PCNA complex as a target for p21WAF1. Science 1997; 277(5334):1996–2000.PubMedCrossRefGoogle Scholar
  54. 54.
    Iida T, Suetake I, Tajima S et al. PCNA clamp facilitates action of DNA cytosine methyltransferase 1 on hemimethylated DNA. Genes Cells 2002; 10:997–1007.CrossRefGoogle Scholar
  55. 55.
    Rountree MR, Bachman KE, Baylin SB. DNMT1 binds HDAC2 and a new co-repressor, DMAP1, to form a complex at replication foci. Nat Genet 2000; 25(3):269–77.PubMedCrossRefGoogle Scholar
  56. 56.
    Pradhan S, Kim GD. The retinoblastoma gene product interacts with maintenance human DNA (cytosine-5) methyltransferase and modulates its activity. EMBO J 2002; 21(4):779–88.PubMedCrossRefGoogle Scholar
  57. 57.
    Robertson KD, Ait-Si-Ali S, Yokochi T et al. DNMT1 forms a complex with Rb, E2F1 and HDAC1 and represses transcription from E2F-responsive promoters. Nat Genet 2000; 25(3):338–42.PubMedCrossRefGoogle Scholar
  58. 58.
    Fatemi M, Hermann A, Gowher H et al. Dnmt3a and Dnmt1 functionally cooperate during de novo methylation of DNA. Eur J Biochem 2002; 269(20):4981–4.PubMedCrossRefGoogle Scholar
  59. 59.
    Belinsky SA, Palmisano WA, Gilliland FD et al. Aberrant promoter methylation in bronchial epithelium and sputum from current and former smokers. Cancer Res 2002; 62(8):2370–7.PubMedGoogle Scholar
  60. 60.
    Plumb JA, Strathdee G, Sludden J et al. Reversal of drug resistance in human tumor xenografts by 2′-deoxy-5-azacytidine-induced demethylation of the hMLH1 gene promoter. Cancer Res 2000;60(21):6039–44.PubMedGoogle Scholar
  61. 61.
    Yoshikawa H, Matsubara K, Qian GS et al. SOCS-1, a negative regulator of the JAK/STAT pathway, is silenced by methylation in human hepatocellular carcinoma and shows growth-suppression activity. Nat Genet 2001; 28(1):29–35.PubMedCrossRefGoogle Scholar
  62. 62.
    Niitsu N, Hayashi Y, Sugita K et al. Sensitization by 5-aza-2′-deoxycytidine of leukaemia cells with MLL abnormalities to induction of differentiation by all-trans retinoic acid and 1alpha,25-dihydroxyvitamin D3. Br J Haematol 2001; 112(2):315–26.PubMedCrossRefGoogle Scholar
  63. 63.
    Harada K, Toyooka S, Shivapurkar N et al. Deregulation of caspase 8 and 10 expression in pediatric tumors and cell lines. Cancer Res 2002; 62(20):5897–901.PubMedGoogle Scholar
  64. 64.
    Esteller M, Corn PG, Baylin SB et al. A gene hypermethylation profile of human cancer. Cancer Res 2001; 61(8):3225–9.PubMedGoogle Scholar
  65. 65.
    Dammann R, Li C, Yoon JH et al. Epigenetic inactivation of a RAS association domain family protein from the lung tumour suppressor locus 3p21.3. Nat Genet 2000; 25(3):315–9.PubMedCrossRefGoogle Scholar
  66. 66.
    Burbee DG, Forgacs E, Zochbauer-Muller S et al. Epigenetic inactivation of RASSF1A in lung and breast cancers and malignant phenotype suppression. J Natl Cancer Inst 2001; 93(9):691–9.PubMedCrossRefGoogle Scholar
  67. 67.
    Serrano A, Tanzarella S, Lionello I et al. Rexpression of HLA class I antigens and restoration of antigen-specific CTL response in melanoma cells following 5-aza-2′-deoxycytidine treatment. Int J Cancer 2001; 94(2):243–51.PubMedCrossRefGoogle Scholar
  68. 68.
    Johnstone RW. Histone-deacetylase inhibitors: novel drugs for the treatment of cancer. Nat Rev Drug Discov 2002; 1(4):287–99.PubMedCrossRefGoogle Scholar
  69. 69.
    Jenuwein T, Allis CD. Translating the histone code. Science 2001; 293(5532):1074–80.PubMedCrossRefGoogle Scholar
  70. 70.
    Abele R, Clavel M, Dodion P et al. The EORTC Early Clinical Trials Cooperative Group experience with 5-aza-2′-deoxycytidine (NSC 127716) in patients with colo-rectal, head and neck, renal carcinomas and malignant melanomas. Eur J Cancer Clin Oncol 1987; 12:1921–4.CrossRefGoogle Scholar
  71. 71.
    Momparler RL, Bouffard DY, Momparler LF et al. Pilot phase I–II study on 5-aza-2′-deoxycytidine (Decitabine) in patients with metastatic lung cancer. Anticancer Drugs 1997; 4:358–68.CrossRefGoogle Scholar
  72. 72.
    Thibault A, Figg WD, Bergan RC et al. A phase II study of 5-aza-2′deoxycytidine (decitabine) in hormone independent metastatic (D2) prostate cancer. Tumori 1998; 1:87–9.Google Scholar
  73. 73.
    Schwartsmann G, Schunemann H, Gorini CN et al. A phase I trial of cisplatin plus decitabine, a new DNA-hypomethylating agent, in patients with advanced solid tumors and a follow-up early phase II evaluation in patients with inoperable non-small cell lung cancer. Invest New Drugs 2000;1:83–91.CrossRefGoogle Scholar
  74. 74.
    Silverman LR, Demakos EP, Peterson B et al. The CALGB, Chicago, IL. A randomized controlled trial of subcutaneous azacitidine (AZA C) in patients with the myelodysplastic syndrome (MDS). A study of the cancer and leukemia GROUP B (CALGB). Proc ASCO 1998.Google Scholar
  75. 75.
    Santini V, Kantarjian HM, Issa JP. Changes in DNA Methylation in Neoplasia: Pathophysiology and Therapeutic Implications. Ann Intern Med 2001; 134:573–86.PubMedGoogle Scholar
  76. 76.
    Wijermans P, Lubbert M, Verhoef G et al. 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 2000; 5:956–62.Google Scholar
  77. 77.
    Aoki E et al. Methylation status of the p15INK4B gene in hematopoietic progenitors and peripheral blood cells in myelodysplastic syndromes. Leukemia 2000; 14:586–93.PubMedCrossRefGoogle Scholar
  78. 78.
    Quesnel B, Fenaux P: p15INK4b gene methylation and myelodysplastic syndromes. Leuk Lymphoma 1999; 35:437–43.PubMedCrossRefGoogle Scholar
  79. 79.
    Pinto A, Zagonel V. 5-Aza-2′-deoxycytidine (Decitabine) and 5-azacytidine in the treatment of acute myeloid leukemias and myelodysplastic syndromes: past, present and future trends. Leukemia 1993; 7(Suppl 1):51–60.PubMedGoogle Scholar
  80. 80.
    Yuen A, Halsey J, Fisher G et al. Phase I/II Trial of ISIS 3521, an Antisense Inhibitor of PKC-Alpha, with Carboplatin and Paclitaxel in Non-Small Cell Lung Cancer. Proc ASCO 2001; 20:1234.Google Scholar
  81. 81.
    Morris MJ et al. Clin Phase I trial of BCL-2 antisense oligonucleotide (G3139) administered by continuous intravenous infusion in patients with advanced cancer. Cancer Res 2002; 8(3):679–83.Google Scholar
  82. 82.
    Stewart DJ, Donehower R C, Eisenhauer EA et al. Phase I study of the DNA methyltransferase 1 inhibitor MG98 administered twice weekly as a two hour intravenous infusion. Ann Onc 2003; 14:766–774.CrossRefGoogle Scholar
  83. 83.
    Davis AJ, Gelmon KA, Siu LL et al. Phase I and Pharmacologic Study of the Human DNA Methyltransferase Antisense Oligodeoxynucleotide MG98 Given as a 21-Day Continuous Infusion Every 4 Weeks. Invest New Drugs 2003; 21:85–97.PubMedCrossRefGoogle Scholar

Copyright information

© and Kluwer Academic/Plenum Publishers 2005

Authors and Affiliations

  • Gregory K. Reid
    • 1
  • A. Robert MacLeod
    • 2
  1. 1.Department of Clinical ResearchMethylGene Inc.MontrealCanada
  2. 2.Department of Molecular BiologyMethylGene Inc.MontrealCanada

Personalised recommendations