Androgen receptor-interacting protein HSPBAP1 facilitates growth of prostate cancer cells in androgen-deficient conditions
Khalid Saeed
Institute for Molecular Medicine Finland, FIMM, University of Helsinki, Helsinki, Finland
Conflict of interests: Nothing to report
Search for more papers by this authorPäivi Östling
Institute for Molecular Medicine Finland, FIMM, University of Helsinki, Helsinki, Finland
Search for more papers by this authorMari Björkman
Medical Biotechnology, VTT Technical Research Centre, Turku, Finland
Search for more papers by this authorTuomas Mirtti
Institute for Molecular Medicine Finland, FIMM, University of Helsinki, Helsinki, Finland
Department of Pathology, Haartman Institute, University of Helsinki and HUSLAB, Helsinki University Central Hospital, Helsinki, Finland
Search for more papers by this authorKalle Alanen
Department of Pathology, Turku University Central Hospital, Turku, Finland
Search for more papers by this authorTiina Vesterinen
Institute for Molecular Medicine Finland, FIMM, University of Helsinki, Helsinki, Finland
Search for more papers by this authorAnna Sankila
Department of Pathology, Haartman Institute, University of Helsinki and HUSLAB, Helsinki University Central Hospital, Helsinki, Finland
Search for more papers by this authorJohan Lundin
Institute for Molecular Medicine Finland, FIMM, University of Helsinki, Helsinki, Finland
Search for more papers by this authorMikael Lundin
Institute for Molecular Medicine Finland, FIMM, University of Helsinki, Helsinki, Finland
Search for more papers by this authorAntti Rannikko
Department of Urology, Helsinki University Central Hospital, Finland
Search for more papers by this authorStig Nordling
Department of Pathology, Haartman Institute, University of Helsinki, Finland
Search for more papers by this authorJohn-Patrick Mpindi
Institute for Molecular Medicine Finland, FIMM, University of Helsinki, Helsinki, Finland
Search for more papers by this authorPekka Kohonen
Medical Biotechnology, VTT Technical Research Centre, Turku, Finland
Search for more papers by this authorKristiina Iljin
Medical Biotechnology, VTT Technical Research Centre, Turku, Finland
Turku Centre for Biotechnology, University of Turku, Turku, Finland
Search for more papers by this authorCorresponding Author
Olli Kallioniemi
Institute for Molecular Medicine Finland, FIMM, University of Helsinki, Helsinki, Finland
Correspondence to: Olli Kallioniemi, MD, PhD, Institute for Molecular Medicine Finland, FIMM, University of Helsinki, Tukholmankatu 8, FIN-00290 Helsinki, Finland, Tel.: +358-50-546–8790, E-mail: [email protected]Search for more papers by this authorJuha K. Rantala
Medical Biotechnology, VTT Technical Research Centre, Turku, Finland
Misvik Biology Corporation, Itäinen pitkäkatu 4 B, FI-20520, Turku, Finland
Search for more papers by this authorKhalid Saeed
Institute for Molecular Medicine Finland, FIMM, University of Helsinki, Helsinki, Finland
Conflict of interests: Nothing to report
Search for more papers by this authorPäivi Östling
Institute for Molecular Medicine Finland, FIMM, University of Helsinki, Helsinki, Finland
Search for more papers by this authorMari Björkman
Medical Biotechnology, VTT Technical Research Centre, Turku, Finland
Search for more papers by this authorTuomas Mirtti
Institute for Molecular Medicine Finland, FIMM, University of Helsinki, Helsinki, Finland
Department of Pathology, Haartman Institute, University of Helsinki and HUSLAB, Helsinki University Central Hospital, Helsinki, Finland
Search for more papers by this authorKalle Alanen
Department of Pathology, Turku University Central Hospital, Turku, Finland
Search for more papers by this authorTiina Vesterinen
Institute for Molecular Medicine Finland, FIMM, University of Helsinki, Helsinki, Finland
Search for more papers by this authorAnna Sankila
Department of Pathology, Haartman Institute, University of Helsinki and HUSLAB, Helsinki University Central Hospital, Helsinki, Finland
Search for more papers by this authorJohan Lundin
Institute for Molecular Medicine Finland, FIMM, University of Helsinki, Helsinki, Finland
Search for more papers by this authorMikael Lundin
Institute for Molecular Medicine Finland, FIMM, University of Helsinki, Helsinki, Finland
Search for more papers by this authorAntti Rannikko
Department of Urology, Helsinki University Central Hospital, Finland
Search for more papers by this authorStig Nordling
Department of Pathology, Haartman Institute, University of Helsinki, Finland
Search for more papers by this authorJohn-Patrick Mpindi
Institute for Molecular Medicine Finland, FIMM, University of Helsinki, Helsinki, Finland
Search for more papers by this authorPekka Kohonen
Medical Biotechnology, VTT Technical Research Centre, Turku, Finland
Search for more papers by this authorKristiina Iljin
Medical Biotechnology, VTT Technical Research Centre, Turku, Finland
Turku Centre for Biotechnology, University of Turku, Turku, Finland
Search for more papers by this authorCorresponding Author
Olli Kallioniemi
Institute for Molecular Medicine Finland, FIMM, University of Helsinki, Helsinki, Finland
Correspondence to: Olli Kallioniemi, MD, PhD, Institute for Molecular Medicine Finland, FIMM, University of Helsinki, Tukholmankatu 8, FIN-00290 Helsinki, Finland, Tel.: +358-50-546–8790, E-mail: [email protected]Search for more papers by this authorJuha K. Rantala
Medical Biotechnology, VTT Technical Research Centre, Turku, Finland
Misvik Biology Corporation, Itäinen pitkäkatu 4 B, FI-20520, Turku, Finland
Search for more papers by this authorAbstract
Hormonal therapies targeting androgen receptor (AR) are effective in prostate cancer (PCa), but often the cancers progress to fatal castrate-resistant disease. Improved understanding of the cellular events during androgen deprivation would help to identify survival and stress pathways whose inhibition could synergize with androgen deprivation. Toward this aim, we performed an RNAi screen on 2,068 genes, including kinases, phosphatases, epigenetic enzymes and other druggable gene targets. High-content cell spot microarray (CSMA) screen was performed in VCaP cells in the presence and absence of androgens with detection of Ki67 and cleaved ADP-ribose polymerase (cPARP) as assays for cell proliferation and apoptosis. Thirty-nine candidate genes were identified, whose silencing inhibited proliferation or induced apoptosis of VCaP cells exclusively under androgen-deprived conditions. One of the candidates, HSPB (heat shock 27 kDa)-associated protein 1 (HSPBAP1), was confirmed to be highly expressed in tumor samples and its mRNA expression levels increased with the Gleason grade. We found that strong HSPBAP1 immunohistochemical staining (IHC) was associated with shorter disease-specific survival of PCa patients compared with negative to moderate staining. Furthermore, we demonstrate that HSPBAP1 interacts with AR in the nucleus of PCa cells specifically during androgen-deprived conditions, occupies chromatin at PSA/klk3 and TMPRSS2/tmprss2 enhancers and regulates their expression. In conclusion, we suggest that HSPBAP1 aids in sustaining cell viability by maintaining AR signaling during androgen-deprived conditions.
Abstract
What's new?
What happens on a cellular level when androgen receptor is blocked in prostate cancer? Cutting off androgen to the tumor alleviates the disease for a while, but eventually androgen receptor levels bounce back and the cancer resurges. How? In this paper, the authors searched for genes that helped the cancer survive despite the lack of androgen. They zeroed in on one gene, HSPBAP1, that correlates with poor survival. Without HSPBAP1, prostate cancer cells could no longer express androgen-receptor target genes. This protein interacts with androgen receptor in the nucleus and appears to maintain AR-signaling in the absence of androgen.
Supporting Information
Additional Supporting Information may be found in the online version of this article.
| Filename | Description |
|---|---|
| ijc29303-sup-0001-suppinfofig01.tif35.4 MB |
Supplementary Information |
| ijc29303-sup-0002-suppinfofig02.tif21.1 MB |
Supplementary Information |
| ijc29303-sup-0003-suppinfofig03.tif26.9 MB |
Supplementary Information |
| ijc29303-sup-0004-suppinfofig04.tif16.7 MB |
Supplementary Information |
| ijc29303-sup-0005-suppinfosup01.doc81.5 KB |
Supplementary Information |
| ijc29303-sup-0006-suppinfotab01.doc16 KB |
Supplementary Information |
| ijc29303-sup-0007-suppinfotab02.doc15.5 KB |
Supplementary Information |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
References
- 1 Heinlein CA, Chang C. Androgen receptor in prostate cancer. Endocr Rev 2004; 25: 276–308.
- 2 Culig Z, Comuzzi B, Steiner H, et al. Expression and function of androgen receptor coactivators in prostate cancer. J Steroid Biochem Mol Biol 2004; 92: 265–71.
- 3 Shi XB, Xue L, Zou JX, et al. Prolonged androgen receptor loading onto chromatin and the efficient recruitment of p160 coactivators contribute to androgen-independent growth of prostate cancer cells. Prostate 2008; 68: 1816–26.
- 4 Xu L, Glass CK, Rosenfeld MG. Coactivator and corepressor complexes in nuclear receptor function. Curr Opin Genet Dev 1999; 9: 140–7.
- 5 Bonkhoff H, Berges R. From pathogenesis to prevention of castration resistant prostate cancer. Prostate 2010; 70: 100–12.
- 6 Montgomery RB, Mostaghel EA, Vessella R, et al. Maintenance of intratumoral androgens in metastatic prostate cancer: a mechanism for castration-resistant tumor growth. Cancer Res 2008; 68: 4447–54.
- 7 Chen CD, Welsbie DS, Tran C, et al. Molecular determinants of resistance to antiandrogen therapy. Nat Med 2004; 10: 33–9.
- 8 Buchanan G, Greenberg NM, Scher HI, et al. Collocation of androgen receptor gene mutations in prostate cancer. Clin Cancer Res 2001; 7: 1273–81.
- 9 Dehm SM, Schmidt LJ, Heemers HV, et al. Splicing of a novel androgen receptor exon generates a constitutively active androgen receptor that mediates prostate cancer therapy resistance. Cancer Res 2008; 68: 5469–77.
- 10 Han G, Buchanan G, Ittmann M, et al. Mutation of the androgen receptor causes oncogenic transformation of the prostate. Proc Natl Acad Sci USA 2005; 102: 1151–6.
- 11 Visakorpi T, Hyytinen E, Koivisto P, et al. In vivo amplification of the androgen receptor gene and progression of human prostate cancer. Nat Genet 1995; 9: 401–6.
- 12 Colombel M, Symmans F, Gil S, et al. Detection of the apoptosis-suppressing oncoprotein bc1–2 in hormone-refractory human prostate cancers. Am J Pathol 1993; 143: 390–400.
- 13 Noble RL. Hormonal control of growth and progression in tumors of Nb rats and a theory of action. Cancer Res 1977; 37: 82–94.
- 14 Rantala JK, Edgren H, Lehtinen L, et al. Integrative functional genomics analysis of sustained polyploidy phenotypes in breast cancer cells identifies an oncogenic profile for GINS2. Neoplasia 2010; 12: 877–88.
- 15 Rantala JK, Makela R, Aaltola AR, et al. A cell spot microarray method for production of high density siRNA transfection microarrays. BMC Genomics 2011; 12: 162.
- 16 Korenchuk S, Lehr JE, MClean L, et al. VCaP, a cell-based model system of human prostate cancer. In Vivo 2001; 15: 163–8.
- 17 Jiang M, Ma Y, Cheng H, et al. Molecular cloning and characterization of a novel human gene (HSPBAP1) from human fetal brain. Cytogenet Cell Genet 2001; 95: 48–51.
- 18 Hong FX, Breitling R, McEntee CW, et al. RankProd: a bioconductor package for detecting differentially expressed genes in meta-analysis. Bioinformatics 2006; 22: 2825–7.
- 19 Norris JD, Chang CY, Wittmann BM, et al. The homeodomain protein HOXB13 regulates the cellular response to androgens. Mol Cell 2009; 36: 405–16.
- 20 Wang H, Zhang C, Rorick A, et al. CCI-779 inhibits cell-cycle G2-M progression and invasion of castration-resistant prostate cancer via attenuation of UBE2C transcription and mRNA stability. Cancer Res 2011; 71: 4866–76.
- 21 Wang Q, Carroll JS, Brown M. Spatial and temporal recruitment of androgen receptor and its coactivators involves chromosomal looping and polymerase tracking. Mol Cell 2005; 19: 631–42.
- 22 Gupta S, Iljin K, Sara H, et al. FZD4 as a mediator of ERG oncogene-induced WNT signaling and epithelial-to-mesenchymal transition in human prostate cancer cells. Cancer Res 2010; 70: 6735–45.
- 23 Soderberg O, Leuchowius KJ, Gullberg M, et al. Characterizing proteins and their interactions in cells and tissues using the in situ proximity ligation assay. Methods 2008; 45: 227–32.
- 24 Reddy TE, Pauli F, Sprouse RO, et al. Genomic determination of the glucocorticoid response reveals unexpected mechanisms of gene regulation. Genome Res 2009; 19: 2163–71.
- 25 Makkonen H, Kauhanen M, Jaaskelainen T, et al. Androgen receptor amplification is reflected in the transcriptional responses of vertebral-cancer of the prostate cells. Mol Cell Endocrinol 2011; 331: 57–65.
- 26 Tomlins SA, Rhodes DR, Perner S, et al. Recurrent fusion of TMPRSS2 and ETS transcription factor genes in prostate cancer. Science 2005; 310: 644–8.
- 27 Rainey WE, Nakamura Y. Regulation of the adrenal androgen biosynthesis. J Steroid Biochem Mol Biol 2008; 108: 281–6.
- 28 Zou JX, Guo L, Revenko AS, et al. Androgen-induced coactivator ANCCA mediates specific androgen receptor signaling in prostate cancer. Cancer Res 2009; 69: 3339–46.
- 29 Meng J, Holdcraft RW, Shima JE, et al. Androgens regulate the permeability of the blood-testis barrier. Proc Natl Acad Sci USA 2005; 102: 16696–700.
- 30 Lubik AA, Gunter JH, Hendy SC, et al. Insulin increases de novo steroidogenesis in prostate cancer cells. Cancer Res 2011; 71: 5754–64.
- 31 Wolf SS, Patchev VK, Obendorf M. A novel variant of the putative demethylase gene, s-JMJD1C, is a coactivator of the AR. Arch Biochem Biophys 2007; 460: 56–66.
- 32 Matzkin ME, Gonzalez-Calvar SI, Mayerhofer A, et al. Testosterone induction of prostaglandin-endoperoxide synthase 2 expression and prostaglandin F(2alpha) production in hamster Leydig cells. Reproduction 2009; 138: 163–75.
- 33 Li J, Ding Z, Wang Z, et al. Androgen regulation of 5alpha-reductase isoenzymes in prostate cancer: implications for prostate cancer prevention. PLoS One 2011; 6: e28840.
- 34 Ting HJ, Yeh S, Nishimura K, et al. Supervillin associates with androgen receptor and modulates its transcriptional activity. Proc Natl Acad Sci USA 2002; 99: 661–6.
- 35 Ghayad SE, Vendrell JA, Bieche I, et al. Identification of TACC1, NOV, and PTTG1 as new candidate genes associated with endocrine therapy resistance in breast cancer. J Mol Endocrinol 2009; 42: 87–103.
- 36 Cai Y, Lee YF, Li G, et al. A new prostate cancer therapeutic approach: combination of androgen ablation with COX-2 inhibitor. Int J Cancer 2008; 123: 195–201.
- 37 Taylor BS, Schultz N, Hieronymus H, et al. Integrative genomic profiling of human prostate cancer. Cancer Cell 2010; 18: 11–22.
- 38 Cerami E, Gao J, Dogrusoz U, et al. The cBio Cancer Genomics Portal: an open platform for exploring multidimensional cancer genomics data (vol 2, pg 401, 2012). Cancer Discov 2012; 2: 960.
- 39 Sahu B, Laakso M, Ovaska K, et al. Dual role of FoxA1 in androgen receptor binding to chromatin, androgen signalling and prostate cancer. EMBO J 2011; 30: 3962–76.
- 40 Bodmer D, Schepens M, Eleveld MJ, et al. Disruption of a novel gene, DIRC3, and expression of DIRC3-HSPBAP1 fusion transcripts in a case of familial renal cell cancer and t(2;3)(q35;q21). Genes Chromosomes Cancer 2003; 38: 107–16.
- 41 Bjorkman M, Rantala J, Nees M, et al. Epigenetics of prostate cancer and the prospect of identification of novel drug targets by RNAi screening of epigenetic enzymes. Epigenomics 2010; 2: 683–9.
- 42 Wu L, Runkle C, Jin HJ, et al. CCN3/NOV gene expression in human prostate cancer is directly suppressed by the androgen receptor. Oncogene 2014; 33: 504–13.
- 43 Yu J, Mani RS, Cao Q, et al. An integrated network of androgen receptor, polycomb, and TMPRSS2-ERG gene fusions in prostate cancer progression. Cancer Cell 2010; 17: 443–54.
- 44 Massie CE, Lynch A, Ramos-Montoya A, et al. The androgen receptor fuels prostate cancer by regulating central metabolism and biosynthesis. EMBO J 2011; 30: 2719–33.
- 45 Chung LW. LNCaP human prostate cancer progression model. Urol Oncol 1996; 2: 126–8.
- 46 Rocchi P, Beraldi E, Ettinger S, et al. Increased Hsp27 after androgen ablation facilitates androgen-independent progression in prostate cancer via signal transducers and activators of transcription 3-mediated suppression of apoptosis. Cancer Res 2005; 65: 11083–93.
- 47 Wang HQ, Yang B, Xu CL, et al. Differential phosphoprotein levels and pathway analysis identify the transition mechanism of LNCaP cells into androgen-independent cells. Prostate 2010; 70: 508–17.
- 48 Zoubeidi A, Zardan A, Beraldi E, et al. Cooperative interactions between androgen receptor (AR) and heat-shock protein 27 facilitate AR transcriptional activity. Cancer Res 2007; 67: 10455–65.
Citing Literature
