1139 SPECIFIC CHANGES IN THE DNA METHYLATION PATTERN ENABLE AN EARLY ASSESSMENT OF THE METASTATIC RISK OF PRIMARY BLADDER TUMOURS

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You have accessJournal of UrologyBladder Cancer: Basic Research (III)1 Apr 20131139 SPECIFIC CHANGES IN THE DNA METHYLATION PATTERN ENABLE AN EARLY ASSESSMENT OF THE METASTATIC RISK OF PRIMARY BLADDER TUMOURS Beatrice Stubendorff, Kerstin Wilhelm, Supriya Dubey, Ewa Dudziec, James Catto, Mieczyslaw Gajda, Heiko Wunderlich, Marc-Oliver Grimm, and Kerstin Junker Beatrice StubendorffBeatrice Stubendorff Jena, Germany More articles by this author , Kerstin WilhelmKerstin Wilhelm Jena, Germany More articles by this author , Supriya DubeySupriya Dubey Jena, Germany More articles by this author , Ewa DudziecEwa Dudziec Oxford, 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 Eisenach, Germany More articles by this author , Marc-Oliver GrimmMarc-Oliver Grimm Jena, Germany More articles by this author , and Kerstin JunkerKerstin Junker Homburg, Germany More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2013.02.754AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Adjuvant chemotherapy improves the outcome of bladder cancer patients at risk for the development of metastases. However, there are no parameters available that allow an individual risk assessment. Therefore, identification of reliable prognostic markers for early prediction of tumour spread is required. The aim of this project is to determine whether changes in DNA methylation correlate with the metastasis risk and to identify a specific DNA methylation pattern that provides a reliable tool for prognosis assessment. METHODS In total, 38 muscle-invasive primary bladder tumours were analysed. Mean follow-up was 2 years. Differences in the methylation pattern were detected by CpG Island (CpGI) microarray including 9 metastatic and 14 non-metastatic samples. Cluster analysis enabled selection of differentially methylated genes. Promoter methylation and gene expression were assessed by quantitative analysis of DNA methylation using real-time PCR (n=38) and qRT-PCR (n=12). For in vitro re-expression T24 cells were exposed to DNMT inhibitor 5-aza-dC. Bladder cancer cells RT112 were transfected using siRNA. Proliferation and migration was measured using real-time cell monitoring. Invasion assay was performed using matrigel coated cell culture inserts. RESULTS Microarray analysis resulted in 201 promoter CpGIs with significant differences (p<0.05) in DNA methylation between metastatic and non-metastatic primary bladder tumours. Validation of methylation status showed differences between patient groups for candidate genes KISS1R, SEPT9 (p<0.05) and CSAD (p<0.01). KISS1R expression differs between the patient groups (p=0.03). Correlation between promoter methylation and mRNA expression was confirmed for all 3 genes by treatment of cell line with 5-aza-dC. SEPT9 transcript variant 3 (SEPT9v3) showed a trend to lower expression in metastatic tumours. Knock down of SEPT9v3 has highest influence on cell proliferation (increased by 77% in compared to non-transfected control). Cell migration and invasion are increased by 15% and 39%, respectively. CONCLUSIONS Primary bladder tumours show significant differences in the DNA methylation pattern in correlation to the metastatic potential. KISS1R shows strongest association with the metastatic risk on methylation and expression level. Reduced expression of SEPT9v3 induces cell proliferation, migration and invasion. The combination of candidate genes KISS1R, CSAD, SEPT9 could serve as prognostic marker panel for determination of the metastatic risk of primary bladder tumours. © 2013 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 189Issue 4SApril 2013Page: e465 Advertisement Copyright & Permissions© 2013 by American Urological Association Education and Research, Inc.MetricsAuthor Information Beatrice Stubendorff Jena, Germany More articles by this author Kerstin Wilhelm Jena, Germany More articles by this author Supriya Dubey Jena, Germany More articles by this author Ewa Dudziec Oxford, 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 Eisenach, Germany More articles by this author Marc-Oliver Grimm Jena, Germany More articles by this author Kerstin Junker Homburg, Germany More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...