Home LiteratureArticle Details
PMID: 14729971 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Continuous zebularine treatment effectively sustains demethylation in human bladder cancer cells.

Molecular and cellular biology ·Vol. 24 ·No. 3 ·2004-02-00 ·Pages 1270-8

Cheng JC, Weisenberger DJ, Gonzales FA, Liang G, Xu GL, Hu YG, Marquez VE, Jones PA

Abstract

During tumorigenesis, tumor suppressor and cancer-related genes are commonly silenced by aberrant DNA methylation in their promoter regions. Recently, we reported that zebularine [1-(beta-D-ribofuranosyl)-1,2-dihydropyrimidin-2-one] acts as an inhibitor of DNA methylation and exhibits chemical stability and minimal cytotoxicity both in vitro and in vivo. Here we show that continuous application of zebularine to T24 cells induces and maintains p16 gene expression and sustains demethylation of the 5' region for over 40 days, preventing remethylation. In addition, continuous zebularine treatment effectively and globally demethylated various hypermethylated regions, especially CpG-poor regions. The drug caused a complete depletion of extractable DNA methyltransferase 1 (DNMT1) and partial depletion of DNMT3a and DNMT3b3. Last, sequential treatment with 5-aza-2'-deoxycytidine followed by zebularine hindered the remethylation of the p16 5' region and gene resilencing, suggesting the possible combination use of both drugs as a potential anticancer regimen.

MeSH Terms
Azacitidine/analogs & derivatives,pharmacology Cytidine/analogs & derivatives DNA/drug effects DNA (Cytosine-5-)-Methyltransferase 1 DNA (Cytosine-5-)-Methyltransferases/antagonists & inhibitors,metabolism DNA Methylation/drug effects Decitabine Enzyme Inhibitors/pharmacology Gene Silencing/drug effects Genes, p16/drug effects Humans Pyrimidine Nucleosides/pharmacology Urinary Bladder Neoplasms/drug therapy
Chemicals
Enzyme Inhibitors Pyrimidine Nucleosides Cytidine Decitabine pyrimidin-2-one beta-ribofuranoside DNA DNA (Cytosine-5-)-Methyltransferase 1 DNA (Cytosine-5-)-Methyltransferases DNMT1 protein, human Azacitidine
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Cheng Jonathan C
Department of Urology, USC/Norris Comprehensive Cancer Center, Keck School of Medicine, University of Southern California, Los Angeles, California 90089-9181, USA.
Weisenberger Daniel J
Gonzales Felicidad A
Liang Gangning
Xu Guo-Liang
Hu Ye-Guang
Marquez Victor E
Jones Peter A
References (42)
42 references, click to expand
  1. Differential mRNA expression of the human DNA methyltransferases (DNMTs) 1, 3a and 3b during the G(0)/G(1) to S phase transition in normal and tumor cells.
    Nucleic Acids Res. 2000 May 15;28(10):2108-13 PMID: 10773079
  2. DNA methylation and cancer.
    J Cell Physiol. 2000 May;183(2):145-54 PMID: 10737890
  3. Cooperativity between DNA methyltransferases in the maintenance methylation of repetitive elements.
    Mol Cell Biol. 2002 Jan;22(2):480-91 PMID: 11756544
  4. Murine de novo methyltransferase Dnmt3a demonstrates strand asymmetry and site preference in the methylation of DNA in vitro.
    Mol Cell Biol. 2002 Feb;22(3):704-23 PMID: 11784849
  5. DNMT1 and DNMT3b cooperate to silence genes in human cancer cells.
    Nature. 2002 Apr 4;416(6880):552-6 PMID: 11932749
  6. Cell division is required for de novo methylation of CpG islands in bladder cancer cells.
    Cancer Res. 2002 Apr 15;62(8):2378-84 PMID: 11956100
  7. The fundamental role of epigenetic events in cancer.
    Nat Rev Genet. 2002 Jun;3(6):415-28 PMID: 12042769
  8. DNA methylation inhibitors in the treatment of leukemias, myelodysplastic syndromes and hemoglobinopathies: clinical results and possible mechanisms of action.
    Curr Top Microbiol Immunol. 2000;249:135-64 PMID: 10802943
  9. Zebularine: a novel DNA methylation inhibitor that forms a covalent complex with DNA methyltransferases.
    J Mol Biol. 2002 Aug 23;321(4):591-9 PMID: 12206775
  10. Histone H3-lysine 9 methylation is associated with aberrant gene silencing in cancer cells and is rapidly reversed by 5-aza-2'-deoxycytidine.
    Cancer Res. 2002 Nov 15;62(22):6456-61 PMID: 12438235
  11. DNMT1 is required to maintain CpG methylation and aberrant gene silencing in human cancer cells.
    Nat Genet. 2003 Jan;33(1):61-5 PMID: 12496760
  12. Inhibition of DNA methylation and reactivation of silenced genes by zebularine.
    J Natl Cancer Inst. 2003 Mar 5;95(5):399-409 PMID: 12618505
  13. Epigenomic stress response. Knockdown of DNA methyltransferase 1 triggers an intra-S-phase arrest of DNA replication and induction of stress response genes.
    J Biol Chem. 2003 Apr 25;278(17):14985-95 PMID: 12576480
  14. DNA methyltransferase 3b contributes to oncogenic transformation induced by SV40T antigen and activated Ras.
    Oncogene. 2003 Jul 24;22(30):4723-33 PMID: 12879017
  15. Functional striated muscle cells from non-myoblast precursors following 5-azacytidine treatment.
    Nature. 1977 May 26;267(5609):364-6 PMID: 68440
  16. Isolation, characterization, and properties of a labile hydrolysis product of the antitumor nucleoside, 5-azacytidine.
    J Med Chem. 1978 Feb;21(2):204-8 PMID: 74412
  17. Kinetics of phosphorylation of 5-aza-2'-deoxyycytidine by deoxycytidine kinase.
    Biochem Pharmacol. 1979 Apr 15;28(8):1443-4 PMID: 87202
  18. Phenotypic conversion of cultured mouse embryo cells by aza pyrimidine nucleosides.
    Dev Biol. 1978 Sep;66(1):57-71 PMID: 87352
  19. Cellular differentiation, cytidine analogs and DNA methylation.
    Cell. 1980 May;20(1):85-93 PMID: 6156004
  20. Induction of thymidine kinase in enzyme-deficient Chinese hamster cells.
    Cell. 1982 Jun;29(2):483-92 PMID: 6180833
  21. DNA methylation in 5-aza-2'-deoxycytidine-resistant variants of C3H 10T1/2 C18 cells.
    Mol Cell Biol. 1984 Oct;4(10):2098-102 PMID: 6209556
  22. Covalent bond formation between a DNA-cytosine methyltransferase and DNA containing 5-azacytosine.
    Proc Natl Acad Sci U S A. 1984 Nov;81(22):6993-7 PMID: 6209710
  23. Synthesis of pyrimidin-2-one nucleosides as acid-stable inhibitors of cytidine deaminase.
    J Med Chem. 1986 Aug;29(8):1374-80 PMID: 3735306
  24. Furanose-pyranose isomerization of reduced pyrimidine and cyclic urea ribosides.
    J Med Chem. 1986 Nov;29(11):2351-8 PMID: 3783592
  25. A genomic sequencing protocol that yields a positive display of 5-methylcytosine residues in individual DNA strands.
    Proc Natl Acad Sci U S A. 1992 Mar 1;89(5):1827-31 PMID: 1542678
  26. Potent inhibitors for the deamination of cytosine arabinoside and 5-aza-2'-deoxycytidine by human cytidine deaminase.
    Cancer Chemother Pharmacol. 1992;30(1):7-11 PMID: 1375134
  27. 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 May;7 Suppl 1:51-60 PMID: 7683358
  28. The mechanism of inhibition of DNA (cytosine-5-)-methyltransferases by 5-azacytosine is likely to involve methyl transfer to the inhibitor.
    Biochem J. 1995 Apr 1;307 ( Pt 1):87-92 PMID: 7536414
  29. Oligonucleotides with fluorescent dyes at opposite ends provide a quenched probe system useful for detecting PCR product and nucleic acid hybridization.
    PCR Methods Appl. 1995 Jun;4(6):357-62 PMID: 7580930
  30. Methylation of the 5' CpG island of the p16/CDKN2 tumor suppressor gene in normal and transformed human tissues correlates with gene silencing.
    Cancer Res. 1995 Oct 15;55(20):4531-5 PMID: 7553622
  31. Improved synthesis of zebularine [1-(beta-D-ribofuranosyl)-dihydropyrimidin-2-one] nucleotides as inhibitors of human deoxycytidylate deaminase.
    J Enzyme Inhib. 1995;9(2):147-62 PMID: 8583252
  32. Real time quantitative PCR.
    Genome Res. 1996 Oct;6(10):986-94 PMID: 8908518
  33. Rapid quantitation of methylation differences at specific sites using methylation-sensitive single nucleotide primer extension (Ms-SNuPE).
    Nucleic Acids Res. 1997 Jun 15;25(12):2529-31 PMID: 9171109
  34. Cancer epigenetics comes of age.
    Nat Genet. 1999 Feb;21(2):163-7 PMID: 9988266
  35. Mechanism-based inhibition of C5-cytosine DNA methyltransferases by 2-H pyrimidinone.
    J Mol Biol. 1999 Feb 19;286(2):389-401 PMID: 9973559
  36. The human DNA methyltransferases (DNMTs) 1, 3a and 3b: coordinate mRNA expression in normal tissues and overexpression in tumors.
    Nucleic Acids Res. 1999 Jun 1;27(11):2291-8 PMID: 10325416
  37. CpG island hypermethylation in human colorectal tumors is not associated with DNA methyltransferase overexpression.
    Cancer Res. 1999 May 15;59(10):2302-6 PMID: 10344733
  38. Roles of cell division and gene transcription in the methylation of CpG islands.
    Mol Cell Biol. 1999 Oct;19(10):6690-8 PMID: 10490608
  39. 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 Mar;18(5):956-62 PMID: 10694544
  40. Whole-genome methylation scan in ICF syndrome: hypomethylation of non-satellite DNA repeats D4Z4 and NBL2.
    Hum Mol Genet. 2000 Mar 1;9(4):597-604 PMID: 10699183
  41. DNA methylator and mismatch repair phenotypes are not mutually exclusive in colorectal cancer cell lines.
    Oncogene. 2000 Feb 17;19(7):943-52 PMID: 10702803
  42. Aberrant patterns of DNA methylation, chromatin formation and gene expression in cancer.
    Hum Mol Genet. 2001 Apr;10(7):687-92 PMID: 11257100
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.24.3.1270-1278.2004
Published
2004-02-00
Pages
1270-8
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC321446
Subset
IM
Grants
NCI NIH HHS · T32 CA009659 · United States
NCI NIH HHS · R01 CA082422 · United States
NCI NIH HHS · 1R01 CA 82422 · United States
NCI NIH HHS · 1R01 CA 83867 · United States
NCI NIH HHS · T32 CA09659 · United States
NCI NIH HHS · R01 CA083867 · United States
External Links
PubMed source DOI: 10.1128/MCB.24.3.1270-1278.2004 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2