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Systematic knockdown of epigenetic enzymes identifies a novel histone demethylase PHF8 overexpressed in prostate cancer with an impact on cell proliferation, migration and invasion

Oncogene volume 31, pages 3444–3456 (2012)Cite this article

Abstract

Our understanding of key epigenetic regulators involved in specific biological processes and cancers is still incomplete, despite great progress in genome-wide studies of the epigenome. Here, we carried out a systematic, genome-wide analysis of the functional significance of 615 epigenetic proteins in prostate cancer (PrCa) cells. We used the high-content cell-spot microarray technology and siRNA silencing of PrCa cell lines for functional screening of cell proliferation, survival, androgen receptor (AR) expression, histone methylation and acetylation. Our study highlights subsets of epigenetic enzymes influencing different cancer cell phenotypes. Plant homeo domain (PHD) finger proteins have a key role in cell survival and histone methylation, whereas histone deacetylases were primarily involved in regulating AR expression. In contrast, JumonjiC-domain (JmjC) containing histone lysine demethylases (KDMs) mainly had an impact on cell proliferation. Our results show that the KDMs JARID1B, PHF8, KDM3A, KDM3B and KDM4A were highly expressed in clinical PrCa samples. The PHD-finger protein 8 (PHF8), a transcriptional coactivator with both PHD- and JmjC-domains, was moderately to strongly expressed in 80% of clinical PrCa samples, whereas 76% of normal and benign samples were negative or only showed weak PHF8 expression. Strong PHF8 expression correlated significantly with high Gleason grade and was borderline significant for poor prognosis. The results of functional PHF8 knockdown implicate a role in cell migration and invasion, as shown by cell motility and 3-D invasion assays. Our study suggests that various cellular phenotypes are regulated by distinct subsets of epigenetic enzymes. Proteins interpreting and modifying histone methylation, such as JmjC-domain and particularly PHD-finger proteins like PHF8, are activated in subsets of PrCa's and promote cancer relevant phenotypes.

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Acknowledgements

We thank Professor Tony Kouzarides from The Gurdon Institute, University of Cambridge for valuable suggestions and insights in constructing the epigenetic siRNA library. We thank Daniela Kleine-Kohlbrecher and Professor Kristian Helin from Biotech Research and Innovation Centre (BRIC), University of Copenhagen, Denmark, for insightful discussions, sharing unpublished results and PHF8 over expression construct. We thank Arho Virkki and Elmar Bucher for assistance with gene expression data normalization. This study was supported by grants from Academy of Finland (grant Nr. 111456 to MN, VH), EU-EPITRON (contract # LSHC-CT-2005-518417 to MB, OK), Marie Curie Canceromics (MEXT-CT-2003-2728 to OK), Cancer Organizations Finland and Sigrid Juselius Foundation (JPM, JR).

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  1. J Rantala

    Present address: 4Present address: OHSU Knight Cancer Institute, Oregon Health & Science University, Portland, OR 97239, USA,

  2. O Kallioniemi and M Nees: These authors contributed equally to this work.

Authors and Affiliations

  1. Medical Biotechnology, VTT Technical Research Centre of Finland, and Center for Biotechnology, University of Turku and Åbo Akademi University, Turku, Finland

    M Björkman, P Östling, V Härmä, J Virtanen, J-P Mpindi, J Rantala, E Kaivanto, P Kohonen, O Kallioniemi & M Nees

  2. Institute for Molecular Medicine Finland (FIMM), University of Helsinki, Helsinki, Finland

    P Östling, J-P Mpindi, T Mirtti, T Vesterinen, M Lundin, A Sankila & O Kallioniemi

  3. Department of Urology, HUSLAB, University of Helsinki, Helsinki, Finland

    A Rannikko

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  1. M Björkman
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Björkman, M., Östling, P., Härmä, V. et al. Systematic knockdown of epigenetic enzymes identifies a novel histone demethylase PHF8 overexpressed in prostate cancer with an impact on cell proliferation, migration and invasion. Oncogene 31, 3444–3456 (2012). https://doi.org/10.1038/onc.2011.512

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