Association of TNFRSF10D DNA-methylation with the survival of melanoma patients.

Gudrun Ratzinger,Peter Fritsch,Barbara Wolf,Heidelinde Fiegl,Simone Mitteregger,Regina Berger,Georg Goebel,Georg Weinlich,Bernhard Zelger

doi:10.3390/ijms150711984

Abstract

In this retrospective pilot study, the DNA-methylation status of genes that have been demonstrated to be involved in melanoma carcinogenesis was analyzed in order to identify novel biomarkers for the risk assessment of melanoma patients. We analyzed DNA extracted from punch-biopsies from 68 formalin-fixed paraffin-embedded (FFPE) melanoma specimens. Using MethyLight PCR, we examined 20 genes in specimens from a training set comprising 36 melanoma patients. Selected candidate genes were validated in a test set using FFPE tissue samples from 32 melanoma patients. First, we identified the TNFRSF10D DNA-methylation status (TNFRSF10D methylated vs. unmethylated) as a prognostic marker for overall (p = 0.001) and for relapse-free survival (p = 0.008) in the training set. This finding was confirmed in the independent test set (n = 32; overall survival p = 0.041; relapse-free survival p = 0.012). In a multivariate Cox-regression analysis including all patients, the TNFRSF10D DNA-methylation status remained as the most significant prognostic parameter for overall and relapse-free survival (relative-risk (RR) of death, 4.6 (95% CI: 2.0–11.0; p < 0.001), RR of relapse, 7.2 (95% CI: 2.8–18.3; p < 0.001)). In this study, we demonstrate that TNFRSF10D DNA-methylation analysis of a small tissue-punch from archival FFPE melanoma tissue is a promising approach to provide prognostic information in patients with melanoma.

Highlights

The incidence of melanoma is increasing in white populations worldwide [1]
In the present pilot study, we aimed to explore whether differences in the DNA-methylation pattern from formalin-fixed paraffin-embedded (FFPE) tissues may be useful as a prognostic marker for the further outcome of non-metastatic melanoma patients
Using MethyLight PCR, we analyzed 20 genes that have been demonstrated to be involved in melanoma tumorigenesis

Summary

Introduction

The incidence of melanoma is increasing in white populations worldwide [1]. In advanced disease, median survival is very poor. The five-year risk of death of patients with advanced (Stage IV) disease is about. Clinical experience demonstrates that some patients with thin neoplasms face recurrence, metastases and death after surgical excision, while others with thick melanomas do not. New prognostic markers, such as metallothionines or genetic subtypes defined by gene expression profiling, have been established; additional reliable markers to select patients for early therapy are still lacking [3,4]. Changes in the status of DNA-methylation, a type of epigenetic alteration, are among the most common molecular alterations in human neoplasia, including melanoma [5,6,7,8,9]. We analyzed the DNA-methylation status of the 5' regions of 20 different genes that are involved in melanoma carcinogenesis and that have previously been shown to be aberrantly methylated in melanoma [7,9,10,11,12,13,14,15,16,17] or other human cancers [18]

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Discussion

Conclusion