TY - JOUR
T1 - Hypermethylation of Cyclin D2 is associated with loss of mRNA expression and tumor development in prostate cancer
AU - Henrique, Rui
AU - Costa, Vera Lúcia
AU - Cerveira, Nuno
AU - Carvalho, André Lopes
AU - Hoque, Mohammad Obaidul
AU - Ribeiro, Franclim Ricardo
AU - Oliveira, Jorge
AU - Teixeira, Manuel Rodrigues
AU - Sidransky, David
AU - Jerónimo, Carmen
N1 - Funding Information:
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 Aperfeiçoa-mento 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).
PY - 2006/11
Y1 - 2006/11
N2 - 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.
AB - 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.
KW - Cyclin D2
KW - Epigenetics
KW - Expression
KW - Methylation
KW - Prostate cancer
KW - Quantitative MSP
KW - Quantitative RT-PCR
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UR - http://www.scopus.com/inward/citedby.url?scp=33750435596&partnerID=8YFLogxK
U2 - 10.1007/s00109-006-0099-4
DO - 10.1007/s00109-006-0099-4
M3 - Article
C2 - 17016690
AN - SCOPUS:33750435596
SN - 0946-2716
VL - 84
SP - 911
EP - 918
JO - Journal of Molecular Medicine
JF - Journal of Molecular Medicine
IS - 11
ER -