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Hypermethylation of Cyclin D2 is associated with loss of mRNA expression and tumor development in prostate cancer

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Abstract

D-type cyclins play a pivotal role in cell cycle regulation and their abnormal expression was associated with several human malignancies. To assess Cyclin D2 promoter methylation status and expression levels in prostate tissues, quantitative methylation-specific PCR and quantitative reverse transcription PCR assays were performed in a large series of prostate carcinomas, high-grade prostatic intraepithelial neoplasias (HGPIN), benign prostate hyperplasias (BPH), normal prostate tissue (NPT) samples, and prostate cancer (PCa) cell lines (before and after demethylating treatment). Methylation levels were correlated with mRNA expression levels and key clinicopathologic parameters. Cyclin D2 promoter methylation was found in 117/118 PCa, 38/38 HGPIN, 24/30 BPH, 11/11 NPT, and 4/4 cell lines. Methylation levels were significantly higher in PCa compared with HGPIN, NPT, and BPH (P<0.0001), correlating with tumor stage and Gleason score (r=0.29, P=0.0014; and r=0.32, P=0.0005, respectively). Conversely, Cyclin D2 mRNA levels were significantly lower in PCa (P<0.01) and a significant inverse correlation between Cyclin D2 methylation and expression levels was found in prostatic tissues (r=−0.61, P<0.000001). Demethylating treatment induced a substantial increase in Cyclin D2 mRNA in LNCaP cells whereas decreased levels were observed in DU-145 and PC-3 cells. We concluded that Cyclin D2 promoter methylation downregulates gene transcription and occurs with high frequency at low levels in normal, hyperplastic, and preneoplastic prostate tissues. Conversely, high Cyclin D2 methylation levels characterize invasive prostatic carcinoma, correlating with clinicopathologic features of tumor aggressiveness.

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Acknowledgements

The expert statistical advice of Dr. Maria José Bento is gratefully acknowledged. R. H. and A. L. C. are the recipients of grants from Liga Portuguesa Contra o Cancro—Núcleo Regional do Norte, Portugal, Portugal, and CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior) (BEX 21303-7), Brazil, respectively. V. L. C., M. R. T., and C. J. are supported by grants from Fundação para a Ciência e a Tecnologia [SFRH/BD/23374/2005 and Projecto de Investigação Plurianual do Centro de Investigação do IPO-Porto (03-05)]. This study was also supported by the “Comissão de Fomento da Investigação em Cuidados de Saúde—Ministério da Saúde,” Portugal. This study was supported by the NIH Grant U01CA84986-04 entitled, “Integrated Development of Novel Molecular Markers—The Early Detection Research Network: Biomarkers Developmental Laboratories” (EDRN Grant).

Disclaimer

Under a licensing agreement between Oncomethylome Sciences SA and the Johns Hopkins University, Dr. Sidransky is entitled to a share of royalty received by the University on sales of products described in this article. Dr. Sidransky owns Oncomethylome Sciences, SA stock, which is subject to certain restrictions under University policy. Dr. Sidransky is a paid consultant to Oncomethylome Sciences SA and is a paid member of the company’s Scientific Advisory Board. The term of this arrangement is being managed by the Johns Hopkins University in accordance with its conflict of interest policies. The remaining authors do not declare any conflict of interest.

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Authors and Affiliations

  1. Department of Pathology, Portuguese Oncology Institute, Porto, Rua Dr. Antonio Bernardino de Almeida, Porto, 4200-072, Portugal

    Rui Henrique

  2. Department of Genetics, Portuguese Oncology Institute, Porto, Rua Dr. Antonio Bernardino de Almeida, Porto, 4200-072, Portugal

    Vera Lúcia Costa, Nuno Cerveira, Franclim Ricardo Ribeiro, Manuel Rodrigues Teixeira & Carmen Jerónimo

  3. Department of Urology, Portuguese Oncology Institute, Porto, Rua Dr. Antonio Bernardino de Almeida, Porto, 4200-072, Portugal

    Jorge Oliveira

  4. Department of Pathology and Molecular Immunology, Institute of Biomedical Sciences Abel Salazar, University of Porto, Largo Prof. Abel Salazar 2, Porto, 4099-003, Portugal

    Rui Henrique, Manuel Rodrigues Teixeira & Carmen Jerónimo

  5. Department of Otolaryngology—Head and Neck Surgery, Head and Neck Cancer Research Division, Johns Hopkins University School of Medicine, 818 Ross, 720 Rutland Avenue, Baltimore, MD, 21205, USA

    André Lopes Carvalho, Mohammad Obaidul Hoque & David Sidransky

  6. Fernando Pessoa University School of Health Sciences, Rua Carlos da Maia, 296, Porto, 4200-150, Portugal

    Carmen Jerónimo

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  1. Rui Henrique
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Correspondence to Carmen Jerónimo.

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Henrique, R., Costa, V., Cerveira, N. et al. Hypermethylation of Cyclin D2 is associated with loss of mRNA expression and tumor development in prostate cancer. J Mol Med 84, 911–918 (2006). https://doi.org/10.1007/s00109-006-0099-4

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  • DOI: https://doi.org/10.1007/s00109-006-0099-4

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