869 A SPECIFIC DNA METHYLATION PATTERN DISTINGUISHES BETWEEN METASTATIC AND NON-METASTATIC BLADDER CANCERS

Abstract

You have accessJournal of UrologyBladder Cancer: Basic Research1 Apr 2011869 A SPECIFIC DNA METHYLATION PATTERN DISTINGUISHES BETWEEN METASTATIC AND NON-METASTATIC BLADDER CANCERS Beatrice Stubendorff, Ewa Dudziec, George Bourghol, James Catto, Mieczyslaw Gajda, Heiko Wunderlich, Kerstin Junker, and Marc-Oliver Grimm Beatrice StubendorffBeatrice Stubendorff Jena, Germany More articles by this author , Ewa DudziecEwa Dudziec Sheffield, United Kingdom More articles by this author , George BourgholGeorge Bourghol Sheffield, United Kingdom More articles by this author , James CattoJames Catto Sheffield, United Kingdom More articles by this author , Mieczyslaw GajdaMieczyslaw Gajda Jena, Germany More articles by this author , Heiko WunderlichHeiko Wunderlich Jena, Germany More articles by this author , Kerstin JunkerKerstin Junker Jena, Germany More articles by this author , and Marc-Oliver GrimmMarc-Oliver Grimm Jena, Germany More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2011.02.693AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES The prognosis for patients with metastatic bladder cancer is poor with a 5-year survival rate of less than 6%. Early prediction of the metastatic risk of primary tumours and effective targeted therapies would lead to improvements in prognosis. Currently no parameters exist for an early and individual prognosis assessment for patients with invasive bladder cancer. Epigenetic alterations are well known regulators of tumour development and progression. Since they affect pathways likely to contribute to tumour events they seem to be reliable biomarkers for an early prediction of the metastatic risk of primary tumours. The objective of this project is to determine whether changes in DNA methylation correlate with the development of metastases and to identify a specific DNA methylation pattern that provides a reliable tool for an early and individual prognosis assessment. METHODS Genomic DNA was isolated from 24 fresh frozen bladder tumour tissues (pT2-4) with and without detectable lymph node metastases. Genomic DNA was sonicated to a fragment size of 200 to 600bp. Sonicated DNA was incubated with 5-Methylcytosin antibody for enrichment of methylated fragments (MeDIP). ∼1.6μg of sonicated DNA were retained as input fraction. Input and IP were labelled with Cy3 and Cy5 dUTP nucleotides. Labelled DNA was competitively hybridizes onto CpG island microarrays. Data were visualized in Genomic Workbench 5.0 and analyzed by Microsoft Excel. RESULTS Microarray analyses enabled the detection of aberrant methylation patterns between patients with non-metastatic and metastatic bladder tumours. First results show hypermethylation of promoters of known tumour suppressor genes (RASSF1) as well as genes that contribute to enhanced cell-cell adhesion (EPHB3, ZYX). There are highly significant differences (p<0.01) between metastatic and non-metastatic patient groups. CONCLUSIONS The results support our hypothesis that muscle-invasive primary tumours of the bladder show a specific methylation pattern that correlates with their metastatic risk. Silencing of tumour suppressors as well as cell adhesion molecules by DNA methylation contributes to tumour progression and spread. Identification of affected pathways will lead to improvements in individual prognosis assessment and development of targeted therapies. Further analysis will include identification of additional candidate genes as well as the validation of the methylation status by methylation specific approaches as well as functional cell culture analysis. © 2011 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 185Issue 4SApril 2011Page: e348 Advertisement Copyright & Permissions© 2011 by American Urological Association Education and Research, Inc.MetricsAuthor Information Beatrice Stubendorff Jena, Germany More articles by this author Ewa Dudziec Sheffield, United Kingdom More articles by this author George Bourghol Sheffield, United Kingdom More articles by this author James Catto Sheffield, United Kingdom More articles by this author Mieczyslaw Gajda Jena, Germany More articles by this author Heiko Wunderlich Jena, Germany More articles by this author Kerstin Junker Jena, Germany More articles by this author Marc-Oliver Grimm Jena, Germany More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...