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DNA-Methylierung in der Urindiagnostik und als Prognosemarker beim Urothelkarzinom der Harnblase

DNA methylation on urinalysis and as a prognostic marker in urothelial cancer of the bladder

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Zusammenfassung

Hintergrund

Die Methylierung von DNA von Tumorpatienten wurde als viel versprechender Marker für die Tumordiagnostik und auch als Prognosemarker für Tumorpatienten beschrieben. Die Zielsetzung dieser Arbeit war die Untersuchung von klinischen Proben von Blasentumorpatienten auf DNA-Methylierung zur Untersuchung in der Urindiagnostik und als Prognosemarker.

Material und Methode

Es wurde mikrodisseziiertes Tumor- und Normalgewebe von 105 Patienten untersucht, bei denen aufgrund eines Blasentumors eine transurethrale Resektion durchgeführt wurde. Zur Untersuchung, ob Methylierung im Urin nachgewiesen werden kann, wurden Urinproben von 37 Tumorpatienten und 20 Kontrollen untersucht. Die Methylierungsanalyse wurde mittels einer methylierungsspezifischen quantitativen Real-time-PCR (Methylight) durchgeführt.

Ergebnisse

Verlaufsdaten sind von 95 der 105 Patienten dokumentiert. Insgesamt erlitten 26 Patienten (27.3%) ein Tumorrezidiv. Wir konnten eine Gruppe von Genen identifizieren, die mit einem Tumorrezidiv assoziiert waren (SOCS-1, STAT-1, BCL-2, DAPK, TIMP-3, E-CADHERIN). In der Kaplan-Meier-Analyse und dem Log-rank-Test konnte eine Korrelation zwischen einer TIMP-3-Methylierung und einem längeren rezidivfreien Intervall nachgewiesen werden. Hinsichtlich der Urinanalysen konnte ein Methylierungsmuster identifiziert werden, mit dem eine Sensitivität von 81,1% bei einer Spezifität von 100% (bei einem tumorfreien Kontrollkollektiv) erzielt werden konnte.

Schlussfolgerung

Unsere Ergebnisse untersteichen die Nützlichkeit von Methylierungsuntersuchungen auch beim Harnbasenkarzinom. Unsere Daten implizieren den Nutzen von Methylierungsuntersuchung sowohl als diagnostischer als auch als Prognosemarker.

Abstract

Introduction and objectives

Detection of promoter hypermethylation has been proposed as a promising tool for cancer diagnosis and as a prognostic marker in various cancers. We studied the versatility of DNA methylation for noninvasive diagnosis and as a prognostic marker for non-muscle-invasive bladder carcinoma.

Methods

Tumor specimens were microdissected and DNA was extracted from 105 paraffin-embedded paraffin specimens from patients undergoing transurethral resection for non-muscle-invasive bladder carcinoma. Urine specimens were collected from patients undergoing cystectomy for bladder cancer and from healthy volunteers. Methylation status was assessed with the real-time quantitative methylation-sensitive PCR (MethyLight). We checked a panel of 20 cancer-associated genes (p14ARF, p16 CDKN2A, STAT-1, SOCS-1, DR-3, DR-6, PIG-7, BCL-2, H-TERT, BAX, EDNRB, DAPK, RASSF-1A, FADD, TMS-1, E-CADHERIN, ICAM-1, TIMP-3, MLH-1, COX-2) for DNA methylation.

Results

Follow-up data were available in 95 of 105 patients (91.4%). A tumor recurrence was observed in 26 patients (27.3%). We could identify six genes (SOCS-1, STAT-1, BCL-2, DAPK, TIMP-3, E-cadherin), where methylation was associated with tumor recurrence. In Kaplan-Meier analysis, TIMP-3 showed a significant association with recurrence-free survival. Methylation of TIMP-3 predicted prolonged disease-free interval. Regarding urinalysis we could identify a pattern of methylation markers including DAPK, BCL-2, and H-TERT that yielded a sensitivity of 81.1% with a specificity of 100% in a cancer-free control population

Conclusions

We present data on the clinical usefulness of methylation analysis in bladder carcinoma. Our data confirm that methylation analysis is a promising tool for bladder cancer diagnosis and prognosis.

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Authors and Affiliations

  1. Klinik und Poliklinik für Urologie, Universität Hamburg,Universitätsklinikum Hamburg Eppendorf, Martinistraße 52, 20246, Hamburg, Deutschland

    M.G. Friedrich, M.I. Toma, J.K.H.F. Chun, T. Steuber, L. Budäus, H. Isbarn & H. Huland

  2. Department of Urology, University of Southern California, Norris Comprehensive Cancer Center, Los Angeles, USA

    M.G. Friedrich

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  1. M.G. Friedrich
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  2. M.I. Toma
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  3. J.K.H.F. Chun
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  4. T. Steuber
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  5. L. Budäus
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  6. H. Isbarn
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  7. H. Huland
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Correspondence to M.G. Friedrich.

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Friedrich, M., Toma, M., Chun, J. et al. DNA-Methylierung in der Urindiagnostik und als Prognosemarker beim Urothelkarzinom der Harnblase. Urologe 46, 761–768 (2007). https://doi.org/10.1007/s00120-007-1360-3

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  • DOI: https://doi.org/10.1007/s00120-007-1360-3

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