© The Author 2006. Published by Oxford University Press.
ARTICLE |
Quantitation of Promoter Methylation of Multiple Genes in Urine DNA and Bladder Cancer Detection
Affiliations of authors: Department of Otolaryngology–Head and Neck Surgery (MOH, OT, AC, ER, WHW, DS), Department of Biostatistics/Oncology (MZ, SNG), The Johns Hopkins School of Medicine, Baltimore, MD; The James Buchanan Brady Urological Institute (MS), Department of Pathology (SB, WHW), Johns Hopkins Medical Institutions, Baltimore, MD; OncoMethylome Sciences, Sart-Tilman (Liege), Belgium (WVC)
Correspondence to: David Sidransky MD, Director, Division of Head and Neck Cancer Research, The Johns Hopkins School of Medicine, 818 Ross Research Building, 720 Rutland Ave., Baltimore, MD 21205-2196 (e-mail: dsidrans{at}jhmi.edu).
Background: The noninvasive identification of bladder tumors may improve disease control and prevent disease progression. Aberrant promoter methylation (i.e., hypermethylation) is a major mechanism for silencing tumor suppressor genes and other cancer-associated genes in many human cancers, including bladder cancer. Methods: A quantitative fluorogenic real-time polymerase chain reaction (PCR) assay was used to examine primary tumor DNA and urine sediment DNA from 15 patients with bladder cancer and 25 control subjects for promoter hypermethylation of nine genes (APC, ARF, CDH1, GSTP1, MGMT, CDKN2A, RAR
2, RASSF1A, and TIMP3) to identify potential biomarkers for bladder cancer. We then used these markers to examine urine sediment DNA samples from an additional 160 patients with bladder cancers of various stages and grades and from an additional 69 age-matched control subjects. Data were analyzed on the basis of a prediction model and were internally validated using a jacknife procedure. All statistical tests were two-sided. Results: For all 15 patients with paired DNA samples, the promoter methylation pattern in urine matched that in the primary tumors. Four genes displayed 100% specificity. Of the 175 bladder cancer patients, 121 (69%, 95% confidence interval [CI] = 62% to 76%) displayed promoter methylation in at least one of these genes (CDKN2A, ARF, MGMT, and GSTP1), whereas all control subjects were negative for such methylation (100% specificity, 95% CI = 96% to 100%). A logistic prediction model using the methylation levels of all remaining five genes was developed and internally validated for subjects who were negative on the four-gene panel. This combined, two-stage predictor produced an internally validated ROC curve with an overall sensitivity of 82% (95% CI = 75 % to 87%) and specificity of 96% (95% CI = 90% to 99%). Conclusion: Testing a small panel of genes with the quantitative methylation–specific PCR assay in urine sediment DNA is a powerful noninvasive approach for the detection of bladder cancer. Larger independent confirmatory cohorts with longitudinal follow-up will be required in future studies to define the impact of this technology on early detection, prognosis, and disease monitoring before clinical application.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  D. M.E.I. Hellebrekers, M. H.F.M. Lentjes, S. M. van den Bosch, V. Melotte, K. A.D. Wouters, K. L.J. Daenen, K. M. Smits, Y. Akiyama, Y. Yuasa, S. Sanduleanu, et al. GATA4 and GATA5 are Potential Tumor Suppressors and Biomarkers in Colorectal Cancer Clin. Cancer Res., June 15, 2009; 15(12): 3990 - 3997. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Zhang, K. D. Meyer, and L. Zhang Fetal Exposure to Cocaine Causes Programming of Prkce Gene Repression in the Left Ventricle of Adult Rat Offspring Biol Reprod, March 1, 2009; 80(3): 440 - 448. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. L. Ostrow, H. L. Park, M. O. Hoque, M. S. Kim, J. Liu, P. Argani, W. Westra, W. V. Criekinge, and D. Sidransky Pharmacologic Unmasking of Epigenetically Silenced Genes in Breast Cancer Clin. Cancer Res., February 15, 2009; 15(4): 1184 - 1191. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Brait, S. Begum, A. L. Carvalho, S. Dasgupta, A. L. Vettore, B. Czerniak, O. L. Caballero, W. H. Westra, D. Sidransky, and M. O. Hoque Aberrant Promoter Methylation of Multiple Genes during Pathogenesis of Bladder Cancer Cancer Epidemiol. Biomarkers Prev., October 1, 2008; 17(10): 2786 - 2794. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. O. Hoque, M. S. Kim, K. L. Ostrow, J. Liu, G. B. A. Wisman, H. L. Park, M. L. Poeta, C. Jeronimo, R. Henrique, A. Lendvai, et al. Genome-Wide Promoter Analysis Uncovers Portions of the Cancer Methylome Cancer Res., April 15, 2008; 68(8): 2661 - 2670. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Yamashita, M. S. Kim, H. L. Park, Y. Tokumaru, M. Osada, H. Inoue, M. Mori, and D. Sidransky HOP/OB1/NECC1 Promoter DNA Is Frequently Hypermethylated and Involved in Tumorigenic Ability in Esophageal Squamous Cell Carcinoma Mol. Cancer Res., January 1, 2008; 6(1): 31 - 41. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Yu, T. Zhu, Z. Wang, H. Zhang, Z. Qian, H. Xu, B. Gao, W. Wang, L. Gu, J. Meng, et al. A Novel Set of DNA Methylation Markers in Urine Sediments for Sensitive/Specific Detection of Bladder Cancer Clin. Cancer Res., December 15, 2007; 13(24): 7296 - 7304. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H.-M. Byun, H.-L. Wong, E. A. Birnstein, E. M. Wolff, G. Liang, and A. S. Yang Examination of IGF2 and H19 Loss of Imprinting in Bladder Cancer Cancer Res., November 15, 2007; 67(22): 10753 - 10758. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Wu, Z. Guo, X. Chang, M. S. Kim, J. K. Nagpal, J. Liu, J. M. Maki, K. I. Kivirikko, S. P. Ethier, B. Trink, et al. LOXL1 and LOXL4 Are Epigenetically Silenced and Can Inhibit Ras/Extracellular Signal-Regulated Kinase Signaling Pathway in Human Bladder Cancer Cancer Res., May 1, 2007; 67(9): 4123 - 4129. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Zhang, A. Darwanto, T. A. Linkhart, L. C. Sowers, and L. Zhang Maternal Cocaine Administration Causes an Epigenetic Modification of Protein Kinase C{epsilon} Gene Expression in Fetal Rat Heart Mol. Pharmacol., May 1, 2007; 71(5): 1319 - 1328. [Abstract] [Full Text] [PDF]
|
|