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p21WAF1/CIP1 gene is inactivated in metastatic prostatic cancer cell lines by promoter methylation

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Prostate Cancer and Prostatic Diseases Submit manuscript

Abstract

Introduction:

p21WAF1/CIP1 may act as a tumour suppressor gene (TSG) and loss of the p21WAF1/CIP1 gene has been reported in several solid tumours. The aim of this study was to see whether p21WAF1/CIP1 was expressed in metastatic prostate cancer cell lines and to determine if there was methylation of the p21WAF1/CIP1 promoter.

Method:

PC3, LNCaP and DU145 metastatic prostate cancer cell lines, 1542NP normal prostate, and RD rhabdomyosarcoma cell lines were cultured in the demethylating agent 5-Aza-2 deoxycytidine (5-Aza-CdR). p21WAF1/CIP1 mRNA expression was analysed by RT-PCR. DNA from untreated cell lines was modified with sodium bisulphite and promoter sequencing was performed.

Results:

p21WAF1/CIP1 was expressed at low or undetectable levels in metastatic prostate cancer cell lines but expression was reactivated by treatment with 5-Aza-CdR. Sequence analysis of the promoter region revealed several sites of methylation at the 5′ end of a CpG island in the PC3, LNCaP and DU145 cell line DNA but not in the normal prostate control DNA. Most notably the Sis-inducible element (SEI)-1—a STAT1-binding site, was methylated.

Conclusions:

In this study, we show that p21WAF1/CIP1 expression in metastatic prostate cancer cell lines is enhanced as a result of demethylation of the DNA. Furthermore, several cytosine residues in the promoter region are methylated, including critical binding sites. The inhibition of the STAT1-signalling pathway by methylation of the promoter may inactivate the p21WAF1/CIP1 TSG in prostate cancer.

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Acknowledgements

Simon Bott is kindly sponsored by a BUF/AstraZeneca scholarship.

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Authors and Affiliations

  1. Prostate Cancer Research Centre, Institute of Urology, University College London, London, UK

    S R J Bott, M Arya, R S Kirby & M Williamson

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  1. S R J Bott
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  2. M Arya
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  3. R S Kirby
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  4. M Williamson
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Correspondence to S R J Bott.

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Bott, S., Arya, M., Kirby, R. et al. p21WAF1/CIP1 gene is inactivated in metastatic prostatic cancer cell lines by promoter methylation. Prostate Cancer Prostatic Dis 8, 321–326 (2005). https://doi.org/10.1038/sj.pcan.4500822

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  • DOI: https://doi.org/10.1038/sj.pcan.4500822

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