Research article

Prognostic relevance of Wnt-inhibitory factor-1 (WIF1) and Dickkopf-3 (DKK3) promoter methylation in human breast cancer

Jürgen Veeck^1,5 , Peter J Wild² , Thomas Fuchs³ , Peter J Schüffler³ , Arndt Hartmann⁴ , Ruth Knüchel¹ and Edgar Dahl¹

¹Molecular Oncology Group, Institute of Pathology, University Hospital of the RWTH Aachen, Pauwelsstrasse 30, D-52074 Aachen, Germany

²Institute of Surgical Pathology, University Hospital Zürich, Schmelzbergstrasse 12, CH-8091, Zürich, Switzerland

³Institute for Computational Science and Department of Computer Science, ETH Zürich, Universitätsstrasse 6, CH-8092 Zürich, Switzerland

⁴Department of Pathology, University of Erlangen, Krankenhausstrasse 12, D-91054 Erlangen, Germany

⁵Current address: Cancer Epigenetics and Biology Program (PEBC), Bellvitge Institute for Biomedical Research (ICO-IDIBELL), Hospital Duran i Reynals, Av. Gran Via de L'Hospitalet 199-203, E-08907 Barcelona, Spain

author email corresponding author email

BMC Cancer 2009, 9:217doi:10.1186/1471-2407-9-217

Published:	1 July 2009

Abstract

Background

Secreted Wnt signaling antagonists have recently been described as frequent targets of epigenetic inactivation in human tumor entities. Since gene silencing of certain Wnt antagonists was found to be correlated with adverse patient survival in cancer, we aimed at investigating a potential prognostic impact of the two Wnt antagonizing molecules WIF1 and DKK3 in breast cancer, which are frequently silenced by promoter methylation in this disease.

Methods

WIF1 and DKK3 promoter methylation were assessed by methylation-specific PCR with bisulfite-converted DNA from 19 normal breast tissues and 150 primary breast carcinomas. Promoter methylation was interpreted in a qualitative, binary fashion. Statistical evaluations included two-sided Fisher's exact tests, univariate log-rank tests of Kaplan-Meier curves as well as multivariate Cox regression analyses.

Results

WIF1 and DKK3 promoter methylation were detected in 63.3% (95/150) and 61.3% (92/150) of breast carcinoma samples, respectively. In normal breast tissues, WIF1 methylation was present in 0% (0/19) and DKK3 methylation in 5.3% (1/19) of samples. In breast carcinomas, WIF1 methylation was significantly associated with methylation of DKK3 (p = 0.009). Methylation of either gene was not associated with clinicopathological parameters, except for DKK3 methylation being associated with patient age (p = 0.007). In univariate analysis, WIF1 methylation was not associated with clinical patient outcome. In contrast, DKK3 methylation was a prognostic factor in patient overall survival (OS) and disease-free survival (DFS). Estimated OS rates after 10 years were 54% for patients with DKK3-methylated tumors, in contrast to patients without DKK3 methylation in the tumor, who had a favorable 97% OS after 10 years (p < 0.001). Likewise, DFS at 10 years for patients harboring DKK3 methylation in the tumor was 58%, compared with 78% for patients with unmethylated DKK3 (p = 0.037). Multivariate analyses revealed that DKK3 methylation was an independent prognostic factor predicting poor OS (hazard ratio (HR): 14.4; 95% confidence interval (CI): 1.9–111.6; p = 0.011), and short DFS (HR: 2.5; 95% CI: 1.0–6.0; p = 0.047) in breast cancer.

Conclusion

Although the Wnt antagonist genes WIF1 and DKK3 show a very similar frequency of promoter methylation in human breast cancer, only DKK3 methylation proves as a novel prognostic marker potentially useful in the clinical management of this disease.