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Role of multi-hnRNP nuclear complex in regulation of tumor suppressor ANXA7 in prostate cancer cells

Oncogene volume 29, pages 2457–2466 (2010)Cite this article

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Abstract

Annexin-A7 (ANXA7) tumor suppressor role has been shown in various tumors, and ANXA7 expression has been particularly lost in androgen-resistant prostate cancers. In this study, we studied ANXA7 regulation in normal prostate versus androgen-sensitive and -resistant prostate cancer cells. Deletion mapping analysis showed lowest ANXA7-promoter activities in androgen-sensitive LNCaP prostate cancer cells. Genomatix analysis of ANXA7 promoter identified a cluster of steroid nuclear hormone receptor elements, including V$GREF (V$GRE.02/ARE.02). Gelshift analysis clearly indicated distinct nuclear protein occupancy at this ANXA7-promoter site (−1086/−890) in prostate cancer (LNCaP, DU145, and PC3) versus normal prostate (PrEC) cells. In matrix-assisted laser desorption time-of-flight mass spectrometry-based search for ANXA7 nuclear regulators, we identified several heterogeneous nuclear ribonucleoproteins (hnRNPs) (A1, A2/B1 and K) attached to the steroid-associated ANXA7-promoter site in the androgen-resistant PC3 prostate cancer cells with high ANXA7 gene copy number, but not in PrEC. The hnPNP role in ANXA7 regulation (that was validated by hnRNPA2/B1 antibody interference) resulted in multiple ANXA7 cDNA and protein products in PC3, but not in PrEC. Ingenuity pathways analysis showed plausible molecular paths between ANXA7 and the hnRNP-associated network in prostate cancer progression. Thus, a multi-hnRNP complex can be responsible for aberrant ANXA7 transcription and splicing, thereby affecting ANXA7 expression pattern and tumor suppressor function in prostate cancer.

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Abbreviations

ANXA7:

Annexin A7

AR:

androgen receptor

GR:

glucocorticoid receptor

hnRNP:

heterogeneous nuclear ribonucleoprotein

IPA:

ingenuity pathways analysis

MALDI-TOF MS:

matrix-assisted laser desorption time-of-flight mass spectrometry

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Acknowledgements

We thank Dr Gideon Dreyfuss (University of Pennsylvania School of Medicine) for providing hnRNPA2/B1 antibody. This work was supported by grants funded by the US Department of Defense (DoD).

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Authors and Affiliations

  1. Department of Anatomy, Physiology and Genetics, Institute for Molecular Medicine, Uniformed Services University School of Medicine (USUHS), Bethesda, MD, USA

    Y Torosyan, M Glasman, K Mezhevaya, S Naga, W Huang, C Paweletz, X Leighton, H B Pollard & M Srivastava

  2. Section for Gene Regulation and Bioinformatics, Center for Prostate Disease Research (CPDR), Rockville, MD, USA

    A Dobi

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  1. Y Torosyan
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  2. A Dobi
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Correspondence to M Srivastava.

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Torosyan, Y., Dobi, A., Glasman, M. et al. Role of multi-hnRNP nuclear complex in regulation of tumor suppressor ANXA7 in prostate cancer cells. Oncogene 29, 2457–2466 (2010). https://doi.org/10.1038/onc.2010.2

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