Androgens repress Bcl-2 expression via activation of the retinoblastoma (RB) protein in prostate cancer cells

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Abstract

The oncogene Bcl-2 is upregulated frequently in prostate tumors following androgen ablation therapy, and Bcl-2 overexpression may contribute to the androgen-refractory relapse of the disease. However, the molecular mechanism underlying androgenic regulation of Bcl-2 in prostate cancer cells is understood poorly. In this study, we demonstrated that no androgen response element (ARE) was identified in the androgen-regulated region of the P1 promoter of Bcl-2 gene, whereas, we provided evidence that the androgenic effect is mediated by E2F1 protein through a putative E2F-binding site in the promoter. We further demonstrated that retinoblastoma (RB) protein plays a critical role in androgen regulation of Bcl-2. The phosphorylation levels of RB at serine residues 780 and 795 were decreased in LNCaP cells treated with androgens. Ectopic expression of a constitutively active form of RB inhibited expression of Bcl-2. Knockdown of endogenous RB protein by an Rb small inference RNA (siRNA) induced an increase in Bcl-2 levels. Most importantly, the effect of androgens on Bcl-2 was abolished completely by specific inhibition of RB function with a mutated E1A. Finally, androgen treatment of LNCaP cells upregulated specifically levels of the cyclin-dependent kinase inhibitors (CDKIs) p15INK4B and p27KIP1. Ectopic expression of p15INK4B and/or p27KIP1 inhibited Bcl-2 expression. Knockdown of endogenous p15INK4B or p27KIP1 protein with a pool of siRNAs diminished androgen-induced downregulation of Bcl-2 expression. Therefore, our data indicate that androgens suppress Bcl-2 expression through negatively modulating activities of the E2F site in the Bcl-2 promoter by activating the CDKI-RB axis.

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Abbreviations

AR:

androgen receptor

ARE:

androgen response element

RB:

retinoblastoma protein

CDK:

cyclin-dependent kinase

CDKI:

cyclin-dependent kinase inhibitor

ChIP:

chromatin immunoprecipitation

siRNA:

small interference RNA

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Acknowledgements

We thank Drs LM Boxer, RJ Kelm, JR Nevins, and Z Zacksenhaus for plasmids. This work was supported in part by a grant (DK60920) from the National Institutes of Health and a grant from the TJ Martell Foundation (DJT), and a developmental award of the Mayo Prostate Cancer SPORE (CA91956) funded by the National Cancer Institute (HH).

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Correspondence to Donald J Tindall.

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Keywords

  • androgen receptor
  • Bcl-2
  • RB
  • transcription regulation
  • prostate cancer

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