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A 90 kDa fragment of filamin A promotes Casodex-induced growth inhibition in Casodex-resistant androgen receptor positive C4-2 prostate cancer cells

Oncogene volume 26, pages 6061–6070 (2007)Cite this article

Abstract

Prostate tumors are initially dependent on androgens for growth, but the majority of patients treated with anti-androgen therapy progress to androgen-independence characterized by resistance to such treatment. This study investigates a novel role for filamin A (FlnA), a 280 kDa cytoskeletal protein (consisting of an actin-binding domain (ABD) followed by 24 sequential repeats), in androgen-independent (AI) growth. Full-length FlnA is cleaved to 170 kDa (ABD+FlnA1–15) and 110 kDa fragments (FlnA16–24); the latter is further cleaved to a 90 kDa fragment (repeats 16–23) capable of nuclear translocation and androgen receptor (AR) binding. Here, we demonstrate that in androgen-dependent LNCaP prostate cancer cells, the cleaved 90 kDa fragment is localized to the nucleus, whereas in its AI subline C4–2, FlnA failed to cleave and remained cytoplasmic. Transfection of FlnA16–24 cDNA in C4–2 cells restored expression and nuclear localization of 90 kDa FlnA. Unlike LNCaP, C4–2 cells proliferate in androgen-reduced medium and in the presence of the AR-antagonist Casodex. They also exhibit increased Akt phosphorylation compared to LNCaP, which may contribute to their AI phenotype. Nuclear expression of 90 kDa FlnA in C4–2 cells decreased Akt phosphorylation, prevented proliferation in androgen-reduced medium and restored Casodex sensitivity. This effect was inhibited by constitutive activation of Akt indicating that FlnA restored Casodex sensitivity in C4–2 cells by decreasing Akt phosphorylation. In addition, FlnA-specific siRNA which depleted FlnA levels, but not control siRNA, induced resistance to Casodex in LNCaP cells. Our results demonstrate that expression of nuclear FlnA is necessary for androgen dependence in these cells.

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Acknowledgements

We thank Dr Barry Furr, AstraZeneca, Cheshire, UK, for the gift of bicalutamide (Casodex), Drs Chong Jin Loy and Eu Leong Yong, National University of Singapore, Singapore, for the gifts of pCMV-FlnA(16–24) and pCMV-FlnA(1–15) plasmids; Dr Thomas Franke, Columbia University, New York, NY, USA, for pCMV-6-myr-Akt-HA and pCMV-6-Akt-K179 M plasmids; Dr Bandana Chatterjee, University of Texas Health Science Center at San Antonio (UTHSCSA) for hPSA-luc construct; Dr AO Brinkmann, Erasmus University, The Netherlands, for wild-type AR (pARO), Dr LuZhe Sun, UTHSCSA, for LNAI tumors and cell lines and Drs Clifford G Tepper and Maria Mudryj, University of California, Davis, for CWR22 and 22Rv1 tumors. This work was supported by a Merit award from the Department of Veterans Affairs and award CA109057 from the National Cancer Institute.

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Authors and Affiliations

  1. Department of Urology, School of Medicine, University of California Davis, Sacramento, CA, USA

    Y Wang, R W deVere White & P M Ghosh

  2. Research Services, VA Northern California Health Care System, Mather, CA, USA

    Y Wang & P M Ghosh

  3. University of Texas Health Science Center, San Antonio, TX, USA

    J I Kreisberg, R G Bedolla & M Mikhailova

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  1. Y Wang
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  2. J I Kreisberg
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Correspondence to P M Ghosh.

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Supplementary Information accompanies the paper on the Oncogene Web site (http://www.nature.com/onc).

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Wang, Y., Kreisberg, J., Bedolla, R. et al. A 90 kDa fragment of filamin A promotes Casodex-induced growth inhibition in Casodex-resistant androgen receptor positive C4-2 prostate cancer cells. Oncogene 26, 6061–6070 (2007). https://doi.org/10.1038/sj.onc.1210435

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