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Disease Markers
Volume 30, Issue 4, Pages 151-161
http://dx.doi.org/10.3233/DMA-2011-0790

Absolute Quantitation of DNA Methylation of 28 Candidate Genes in Prostate Cancer Using Pyrosequencing

Nataڑa Vasiljeviš,1 Keqiang Wu,1 Adam R. Brentnall,1 Dae Cheol Kim,1 Mangesh A. Thorat,1 Sakunthala C. Kudahetti,2 Xueying Mao,2 Liyan Xue,2 Yongwei Yu,3 Greg L. Shaw,4 Luis Beltran,4 Yong-Jie Lu,2 Daniel M. Berney,2 Jack Cuzick,1 and Attila T. Lorincz1

1Cancer Research UK Centre for Epidemiology, Mathematics and Statistics, Wolfson Institute of Preventive Medicine, Barts & the London School of Dentistry and Medicine, Queen Mary University of London, London, UK
2Molecular Oncology and Imaging, Institute of Cancer, Barts & the London School of Dentistry and Medicine, Queen Mary University of London, London, UK
3Department of Pathology, Changhai Hospital, The Second Military Medical University, Shanghai, China
4Department of Histopathology, Whipps Cross Hospital, London, UK

Received 17 June 2011; Accepted 17 June 2011

Copyright © 2011 Hindawi Publishing Corporation. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Aberrant DNA methylation plays a pivotal role in carcinogenesis and its mapping is likely to provide biomarkers for improved diagnostic and risk assessment in prostate cancer (PCa). We quantified and compared absolute methylation levels among 28 candidate genes in 48 PCa and 29 benign prostate hyperplasia (BPH) samples using the pyrosequencing (PSQ) method to identify genes with diagnostic and prognostic potential.

RARB, HIN1, BCL2, GSTP1, CCND2, EGFR5, APC, RASSF1A, MDR1, NKX2-5, CDH13, DPYS, PTGS2, EDNRB, MAL, PDLIM4, HLAa, ESR1 and TIG1 were highly methylated in PCa compared to BPH (p < 0.001), while SERPINB5, CDH1, TWIST1, DAPK1, THRB, MCAM, SLIT2, CDKN2a and SFN were not. RARB methylation above 21% completely distinguished PCa from BPH. Separation based on methylation level of SFN, SLIT2 and SERPINB5 distinguished low and high Gleason score cancers, e.g. SFN and SERPINB5 together correctly classified 81% and 77% of high and low Gleason score cancers respectively. Several genes including CDH1 previously reported as methylation markers in PCa were not confirmed in our study. Increasing age was positively associated with gene methylation (p < 0.0001).

Accurate quantitative measurement of gene methylation in PCa appears promising and further validation of genes like RARB, HIN1, BCL2, APC and GSTP1 is warranted for diagnostic potential and SFN, SLIT2 and SERPINB5 for prognostic potential.