© 2003 by Oxford University Press
Journal of the National Cancer Institute, Vol. 95, No. 5, 399-409,
March 5, 2003
© 2003 Oxford University Press
ARTICLE |
Inhibition of DNA Methylation and Reactivation of Silenced Genes by Zebularine
Affiliations of authors: J. C. Cheng, F. A. Gonzales, P. A. Jones, University of Southern California (USC)/Norris Comprehensive Cancer Center and Hospital, Departments of Biochemistry and Molecular Biology and Urology, USC Keck School of Medicine, Los Angeles; C. B. Matsen, E. U. Selker, Institute of Molecular Biology, University of Oregon, Eugene; W. Ye, USC/Norris Comprehensive Cancer Center and Hospital, Department of Preventive Medicine, and USC Keck School of Medicine, Los Angeles; S. Greer, Department of Microbiology and Immunology, Biochemistry and Molecular Biology and Radiation Oncology, University of Miami School of Medicine, Sylvester Cancer Center, Miami, FL; V. E. Marquez, Laboratory of Medicinal Chemistry, Center for Cancer Research, National Cancer Institute at Frederick, Frederick, MD.
Correspondence to: Peter A. Jones, Ph.D., D.Sc., University of Southern California (USC)/Norris Comprehensive Cancer Center and Hospital, Departments of Biochemistry and Molecular Biology and Urology, USC Keck School of Medicine, 1441 Eastlake Ave., Los Angeles, CA 90089-9181 (e-mail: jones_p{at}ccnt.hsc.usc.edu).
Background: Gene silencing by abnormal methylation of promoter regions of regulatory genes is commonly associated with cancer. Silenced tumor suppressor genes are obvious targets for reactivation by methylation inhibitors such as 5-azacytidine (5-Aza-CR) and 5-aza-2'-deoxycytidine (5-Aza-CdR). However, both compounds are chemically unstable and toxic and neither can be given orally. We characterized a new demethylating agent, zebularine [1-(beta-D-ribofuranosyl)-1,2-dihydropyrimidin-2-one], which is a chemically stable cytidine analog. Methods: We tested the ability of zebularine to reactivate a silenced Neurospora crassa gene using a hygromycin gene reactivation assay. We then analyzed the ability of zebularine to inhibit DNA methylation in C3H 10T1/2 Cl8 (10T1/2) mouse embryo cells as assayed by induction of a myogenic phenotype and in T24 human bladder carcinoma cells, using the methylation-sensitive single nucleotide primer extension (Ms-SNuPE) assay. We also evaluated the effects of zebularine (administered orally or intraperitoneally) on growth of EJ6 human bladder carcinoma cells grown in BALB/c nu/nu mice (five mice per group) and the in vivo reactivation of a methylated p16 gene in these cells. All statistical tests were two-sided. Results: In N. crassa, zebularine inhibited DNA methylation and reactivated a gene previously silenced by methylation. Zebularine induced the myogenic phenotype in 10T1/2 cells, which is a phenomenon unique to DNA methylation inhibitors. Zebularine reactivated a silenced p16 gene and demethylated its promoter region in T24 bladder carcinoma cells in vitro and in tumors grown in mice. Zebularine was only slightly cytotoxic to T24 cells in vitro (1 mM zebularine for 48 hours decreased plating efficiency by 17% [95% confidence interval (CI) = 12.8% to 21.2%]) and to tumor-bearing mice (average maximal weight change in mice treated with 1000 mg/kg zebularine = 11% [95% CI = 4% to 19%]). Compared with those in control mice, tumor volumes were statistically significantly reduced in mice treated with high-dose zebularine administered by intraperitoneal injection (P<.001) or by oral gavage (P<.001). Conclusions: Zebularine is a stable DNA demethylating agent and the first drug in its class able to reactivate an epigenetically silenced gene by oral administration.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  W. G. Nelson, A. M. De Marzo, and S. Yegnasubramanian Epigenetic Alterations in Human Prostate Cancers Endocrinology, September 1, 2009; 150(9): 3991 - 4002. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Ellis, P. W. Atadja, and R. W. Johnstone Epigenetics in cancer: Targeting chromatin modifications Mol. Cancer Ther., June 1, 2009; 8(6): 1409 - 1420. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Z. Liu, S. Liu, Z. Xie, R. E. Pavlovicz, J. Wu, P. Chen, J. Aimiuwu, J. Pang, D. Bhasin, P. Neviani, et al. Modulation of DNA Methylation by a Sesquiterpene Lactone Parthenolide J. Pharmacol. Exp. Ther., May 1, 2009; 329(2): 505 - 514. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. B. Yoo, J. C. Chuang, H.-M. Byun, G. Egger, A. S. Yang, L. Dubeau, T. Long, P. W. Laird, V. E. Marquez, and P. A. Jones Long-term Epigenetic Therapy with Oral Zebularine Has Minimal Side Effects and Prevents Intestinal Tumors in Mice Cancer Prevention Research, September 1, 2008; 1(4): 233 - 240. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J.-P. J Issa DNA Methylation as a Therapeutic Target in Cancer Am. Assoc. Cancer Res. Educ. Book, April 12, 2008; 2008(1): 643 - 649. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. B. Yoo, S. Jeong, G. Egger, G. Liang, P. Phiasivongsa, C. Tang, S. Redkar, and P. A. Jones Delivery of 5-Aza-2'-Deoxycytidine to Cells Using Oligodeoxynucleotides Cancer Res., July 1, 2007; 67(13): 6400 - 6408. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Biermann and K. Steger Epigenetics in Male Germ Cells J Androl, July 1, 2007; 28(4): 466 - 480. [Full Text] [PDF]
|
|
|
|
 |
|
 C. S. Zorn, K. J. Wojno, M. T. McCabe, R. Kuefer, J. E. Gschwend, and M. L. Day 5-Aza-2'-Deoxycytidine Delays Androgen-Independent Disease and Improves Survival in the Transgenic Adenocarcinoma of the Mouse Prostate Mouse Model of Prostate Cancer Clin. Cancer Res., April 1, 2007; 13(7): 2136 - 2143. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J.-P. J. Issa DNA Methylation as a Therapeutic Target in Cancer Clin. Cancer Res., March 15, 2007; 13(6): 1634 - 1637. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. P. Mitra, R. H. Datar, and R. J. Cote Molecular Pathways in Invasive Bladder Cancer: New Insights Into Mechanisms, Progression, and Target Identification J. Clin. Oncol., December 10, 2006; 24(35): 5552 - 5564. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. M.E.I. Hellebrekers, K. Castermans, E. Vire, R. P.M. Dings, N. T.H. Hoebers, K. H. Mayo, M. G.A. oude Egbrink, G. Molema, F. Fuks, M. van Engeland, et al. Epigenetic Regulation of Tumor Endothelial Cell Anergy: Silencing of Intercellular Adhesion Molecule-1 by Histone Modifications. Cancer Res., November 15, 2006; 66(22): 10770 - 10777. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. H. Beumer, E. Joseph, M. J. Egorin, R. S. Parker, D. Z. D'argenio, J. M. Covey, and J. L. Eiseman A Mass Balance and Disposition Study of the DNA Methyltransferase Inhibitor Zebularine (NSC 309132) and Three of Its Metabolites in Mice. Clin. Cancer Res., October 1, 2006; 12(19): 5826 - 5833. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Stresemann, B. Brueckner, T. Musch, H. Stopper, and F. Lyko Functional diversity of DNA methyltransferase inhibitors in human cancer cell lines. Cancer Res., March 1, 2006; 66(5): 2794 - 2800. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. M.E.I. Hellebrekers, K.-W. Jair, E. Vire, S. Eguchi, N. T.H. Hoebers, M. F. Fraga, M. Esteller, F. Fuks, S. B. Baylin, M. van Engeland, et al. Angiostatic activity of DNA methyltransferase inhibitors. Mol. Cancer Ther., February 1, 2006; 5(2): 467 - 475. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Herranz, J. Martin-Caballero, M. F. Fraga, J. Ruiz-Cabello, J. M. Flores, M. Desco, V. Marquez, and M. Esteller The novel DNA methylation inhibitor zebularine is effective against the development of murine T-cell lymphoma Blood, February 1, 2006; 107(3): 1174 - 1177. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. H. Lee, S. Yegnasubramanian, X. Lin, and W. G. Nelson Procainamide Is a Specific Inhibitor of DNA Methyltransferase 1 J. Biol. Chem., December 9, 2005; 280(49): 40749 - 40756. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F. Lyko and R. Brown DNA Methyltransferase Inhibitors and the Development of Epigenetic Cancer Therapies J Natl Cancer Inst, October 19, 2005; 97(20): 1498 - 1506. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Balch, P. Yan, T. Craft, S. Young, D. G. Skalnik, T. H-M. Huang, and K. P. Nephew Antimitogenic and chemosensitizing effects of the methylation inhibitor zebularine in ovarian cancer Mol. Cancer Ther., October 1, 2005; 4(10): 1505 - 1514. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. C. Chuang, C. B. Yoo, J. M. Kwan, T. W.H. Li, G. Liang, A. S. Yang, and P. A. Jones Comparison of biological effects of non-nucleoside DNA methylation inhibitors versus 5-aza-2'-deoxycytidine Mol. Cancer Ther., October 1, 2005; 4(10): 1515 - 1520. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Fan, E.-i. Kodama, Y. Koh, M. Nakao, and M. Matsuoka Halogenated Thymidine Analogues Restore the Expression of Silenced Genes without Demethylation Cancer Res., August 1, 2005; 65(15): 6927 - 6933. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Brueckner, R. Garcia Boy, P. Siedlecki, T. Musch, H. C. Kliem, P. Zielenkiewicz, S. Suhai, M. Wiessler, and F. Lyko Epigenetic Reactivation of Tumor Suppressor Genes by a Novel Small-Molecule Inhibitor of Human DNA Methyltransferases Cancer Res., July 15, 2005; 65(14): 6305 - 6311. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Dote, D. Cerna, W. E. Burgan, D. J. Carter, M. A. Cerra, M. G. Hollingshead, K. Camphausen, and P. J. Tofilon Enhancement of In vitro and In vivo Tumor Cell Radiosensitivity by the DNA Methylation Inhibitor Zebularine Clin. Cancer Res., June 15, 2005; 11(12): 4571 - 4579. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. N. Bhalla Epigenetic and Chromatin Modifiers As Targeted Therapy of Hematologic Malignancies J. Clin. Oncol., June 10, 2005; 23(17): 3971 - 3993. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. R. Hodge, B. Peng, J. C. Cherry, E. M. Hurt, S. D. Fox, J. A. Kelley, D. J. Munroe, and W. L. Farrar Interleukin 6 Supports the Maintenance of p53 Tumor Suppressor Gene Promoter Methylation Cancer Res., June 1, 2005; 65(11): 4673 - 4682. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. L. Holleran, R. A. Parise, E. Joseph, J. L. Eiseman, J. M. Covey, E. R. Glaze, A. V. Lyubimov, Y.-F. Chen, D. Z. D'Argenio, and M. J. Egorin Plasma Pharmacokinetics, Oral Bioavailability, and Interspecies Scaling of the DNA Methyltransferase Inhibitor, Zebularine Clin. Cancer Res., May 15, 2005; 11(10): 3862 - 3868. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. J. Cote, P. W. Laird, and R. H. Datar Promoter Hypermethylation: A New Therapeutic Target Emerges in Urothelial Cancer J. Clin. Oncol., May 1, 2005; 23(13): 2879 - 2881. [Full Text] [PDF]
|
|
|
|
|
|
J. W.F. Catto, A.-R. Azzouzi, I. Rehman, K. M. Feeley, S. S. Cross, N. Amira, G. Fromont, M. Sibony, O. Cussenot, M. Meuth, et al. Promoter Hypermethylation Is Associated With Tumor Location, Stage, and Subsequent Progression in Transitional Cell Carcinoma J. Clin. Oncol., May 1, 2005; 23(13): 2903 - 2910. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. W. Laird Cancer epigenetics Hum. Mol. Genet., April 15, 2005; 14(suppl_1): R65 - R76. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. B. Yoo and P. A. Jones DNA Methyltransferase Inhibitors in Cancer Therapy Am. Assoc. Cancer Res. Educ. Book, April 1, 2005; 2005(1): 333 - 337. [Full Text] [PDF]
|
|
|
|
|
|
C. J. Fabian and B. F. Kimler Selective Estrogen-Receptor Modulators for Primary Prevention of Breast Cancer J. Clin. Oncol., March 10, 2005; 23(8): 1644 - 1655. [Full Text] [PDF]
|
|
|
|
|
|
P. M. Das and R. Singal DNA Methylation and Cancer J. Clin. Oncol., November 15, 2004; 22(22): 4632 - 4642. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
W. G. Alleman, R. L. Tabios, G. V. R. Chandramouli, O. N. Aprelikova, C. Torres-Cabala, A. Mendoza, C. Rodgers, N. A. Sopko, W. M. Linehan, and J. R. Vasselli The In vitro and In vivo Effects of Re-Expressing Methylated von Hippel-Lindau Tumor Suppressor Gene in Clear Cell Renal Carcinoma with 5-Aza-2'-deoxycytidine Clin. Cancer Res., October 15, 2004; 10(20): 7011 - 7021. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Gilbert, S. D. Gore, J. G. Herman, and M. A. Carducci The Clinical Application of Targeting Cancer through Histone Acetylation and Hypomethylation Clin. Cancer Res., July 15, 2004; 10(14): 4589 - 4596. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Graziano, F. Arduini, A. Ruzzo, I. Bearzi, B. Humar, H. More, R. Silva, P. Muretto, P. Guilford, E. Testa, et al. Prognostic Analysis of E-Cadherin Gene Promoter Hypermethylation in Patients with Surgically Resected, Node-Positive, Diffuse Gastric Cancer Clin. Cancer Res., April 15, 2004; 10(8): 2784 - 2789. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F. Graziano, F. Arduini, A. Ruzzo, A. Mandolesi, I. Bearzi, R. Silva, P. Muretto, E. Testa, D. Mari, M. Magnani, et al. Combined analysis of E-cadherin gene (CDH1) promoter hypermethylation and E-cadherin protein expression in patients with gastric cancer: implications for treatment with demethylating drugs Ann. Onc., March 1, 2004; 15(3): 489 - 492. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Ghoshal, S. Majumder, J. Datta, T. Motiwala, S. Bai, S. M. Sharma, W. Frankel, and S. T. Jacob Role of Human Ribosomal RNA (rRNA) Promoter Methylation and of Methyl-CpG-binding Protein MBD2 in the Suppression of rRNA Gene Expression J. Biol. Chem., February 20, 2004; 279(8): 6783 - 6793. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. C. Cheng, D. J. Weisenberger, F. A. Gonzales, G. Liang, G.-L. Xu, Y.-G. Hu, V. E. Marquez, and P. A. Jones Continuous Zebularine Treatment Effectively Sustains Demethylation in Human Bladder Cancer Cells Mol. Cell. Biol., February 1, 2004; 24(3): 1270 - 1278. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. F. List, J. Vardiman, J.-P. J. Issa, and T. M. DeWitte Myelodysplastic Syndromes Hematology, January 1, 2004; 2004(1): 297 - 317. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Kopelovich, J. A. Crowell, and J. R. Fay The Epigenome as a Target for Cancer Chemoprevention J Natl Cancer Inst, December 3, 2003; 95(23): 1747 - 1757. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. G. Herman and S. B. Baylin Gene Silencing in Cancer in Association with Promoter Hypermethylation N. Engl. J. Med., November 20, 2003; 349(21): 2042 - 2054. [Full Text] [PDF]
|
|
|
|
|
|
M. Z. Fang, Y. Wang, N. Ai, Z. Hou, Y. Sun, H. Lu, W. Welsh, and C. S. Yang Tea Polyphenol (-)-Epigallocatechin-3-Gallate Inhibits DNA Methyltransferase and Reactivates Methylation-Silenced Genes in Cancer Cell Lines Cancer Res., November 15, 2003; 63(22): 7563 - 7570. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. A. Belinsky, D. M. Klinge, C. A. Stidley, J.-P. Issa, J. G. Herman, T. H. March, and S. B. Baylin Inhibition of DNA Methylation and Histone Deacetylation Prevents Murine Lung Cancer Cancer Res., November 1, 2003; 63(21): 7089 - 7093. [Abstract] [Full Text] [PDF]
|
|