Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

OCT-4: a novel estrogen receptor-α collaborator that promotes tamoxifen resistance in breast cancer cells

Oncogene volume 35, pages 5722–5734 (2016)Cite this article

Subjects

Abstract

Tamoxifen has shown great success in the treatment of breast cancer; however, long-term treatment can lead to acquired tamoxifen (TOT) resistance and relapse. TOT classically antagonizes estradiol (E2) -dependent breast cancer cell growth, but exerts partial agonist/antagonist behavior on gene expression. Although both E2 and TOT treatment of breast cancer cells results in recruitment of the estrogen receptor (ER) to common and distinct genomic sites, the mechanisms and proteins underlying TOT preferential recruitment of the ER remains poorly defined. To this end, we performed in silico motif-enrichment analyses within the ER-binding peaks in response to E2 or TOT, to identify factors that would specifically recruit ER to genomic binding sites in the presence of TOT as compared to E2. Intriguingly, we found Nkx3-1 and Oct-transcription factor homodimer motifs to be enriched in TOT preferential binding sites and confirmed the critical role of Oct-3/4 (aka Oct-4) in directing ER recruitment to TOT preferential genomic binding sites, by chromatin immunoprecipitation (ChIP) analyses. Further investigation revealed Oct-4 expression to be basally repressed by Nkx3-1 in MCF-7 cells and TOT treatment appeared to elevate Nkx3-1 degradation through a p38MAPK-dependent phosphorylation of the E3 ligase, Skp2 at serine-64 residue, as observed by quantitative mass-spectrometry analyses. Consistently, Oct-4 upon induction by phospho-Ser64-Skp2-mediated proteasomal degradation of Nkx3-1, participated in ER transcriptional complexes along with p38MAPK and Skp2 in a tamoxifen-dependent manner leading to TOT-dependent gene activation and cell proliferation of the TOT-resistant MCF-7-tamr breast cancer cells. Notably, Oct-4 levels were highly elevated in MCF-7-tamr cells, and appeared critical for their TOT sensitivity in cell proliferation assays. Furthermore, overexpression of Oct-4 enhanced tumor growth in the presence of tamoxifen in mice in vivo. Collectively, our work presents a novel mechanism for tamoxifen-specific gene activation by ER, secondary to its TOT preferential recruitment to genomic sites by specific activation of Oct-4, a phenomenon that appears to underlie tamoxifen resistance in breast cancer cells and in xenograft tumor models, and could be useful in designing therapeutic interventions to improve treatment outcome.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8

Similar content being viewed by others

Abbreviations

CoIP:

Co-immunoprecipitation

Ub:

ubiquitin

E2:

estradiol

TOT:

tamoxifen

ER:

estrogen receptor

Skp2:

S-phase Kinase-associated Protein-2

p38MAPK:

p38-Mitogen Activated Protein Kinase

NFATc4:

Nuclear Factor Activated T-cell 4

siRNA:

short interfering RNA

Ad:

Adenovirus

ERE:

estrogen response element.

References

  1. Shang Y, Hu X, DiRenzo J, Lazar MA, Brown M . Cofactor dynamics and sufficiency in estrogen receptor-regulated transcription. Cell 2000; 103: 843–852.

    Article  CAS  Google Scholar 

  2. Green KA, Carroll JS . Oestrogen-receptor-mediated transcription and the influence of co-factors and chromatin state. Nat Rev Cancer 2007; 7: 713–722.

    Article  CAS  Google Scholar 

  3. Fullwood MJ, Liu MH, Pan YF, Liu J, Xu H, Mohamed YB et al. An oestrogen-receptor-alpha-bound human chromatin interactome. Nature 2009; 462: 58–64.

    Article  CAS  Google Scholar 

  4. Welboren WJ, Sweep FC, Span PN, Stunnenberg HG . Genomic actions of estrogen receptor alpha: what are the targets and how are they regulated? Endocr Relat Cancer 2009; 16: 1073–1089.

    Article  CAS  Google Scholar 

  5. Katzenellenbogen BS, Katzenellenbogen JA . Biomedicine. Defining the "S" in SERMs. Science 2002; 295: 2380–2381.

    Article  CAS  Google Scholar 

  6. Shang Y, Brown M . Molecular determinants for the tissue specificity of SERMs. Science 2002; 295: 2465–2468.

    Article  CAS  Google Scholar 

  7. Katzenellenbogen BS, Frasor J . Therapeutic targeting in the estrogen receptor hormonal pathway. Semin Oncol 2004; 31: 28–38.

    Article  CAS  Google Scholar 

  8. Brueggemeier RW, Hackett JC, Diaz-Cruz ES . Aromatase inhibitors in the treatment of breast cancer. Endocr Rev 2005; 26: 331–345.

    Article  CAS  Google Scholar 

  9. Jordan VC, O'Malley BW . Selective estrogen-receptor modulators and antihormonal resistance in breast cancer. J Clin Oncol 2007; 25: 5815–5824.

    Article  CAS  Google Scholar 

  10. Jaiyesimi IA, Buzdar AU, Decker DA, Hortobagyi GN . Use of tamoxifen for breast cancer: twenty-eight years later. J Clin Oncol 1995; 13: 513–529.

    Article  CAS  Google Scholar 

  11. Herman ME, Katzenellenbogen BS . Response-specific antiestrogen resistance in a newly characterized MCF-7 human breast cancer cell line resulting from long-term exposure to trans-hydroxytamoxifen. J Steroid Biochem Mol Biol 1996; 59: 121–134.

    Article  CAS  Google Scholar 

  12. Connor CE, Norris JD, Broadwater G, Willson TM, Gottardis MM, Dewhirst MW et al. Circumventing tamoxifen resistance in breast cancers using antiestrogens that induce unique conformational changes in the estrogen receptor. Cancer Res 2001; 61: 2917–2922.

    CAS  PubMed  Google Scholar 

  13. Osborne CK . Tamoxifen in the treatment of breast cancer. N Engl J Med 1998; 339: 1609–1618.

    Article  CAS  Google Scholar 

  14. Bianco S, Gevry N . Endocrine resistance in breast cancer: from cellular signaling pathways to epigenetic mechanisms. Transcription 2012; 3: 165–170.

    Article  Google Scholar 

  15. Huber-Keener KJ, Liu X, Wang Z, Wang Y, Freeman W, Wu S et al. Differential gene expression in tamoxifen-resistant breast cancer cells revealed by a new analytical model of RNA-Seq data. PLoS One 2012; 7: e41333.

    Article  CAS  Google Scholar 

  16. Frasor J, Stossi F, Danes JM, Komm B, Lyttle CR, Katzenellenbogen BS . Selective estrogen receptor modulators: discrimination of agonistic versus antagonistic activities by gene expression profiling in breast cancer cells. Cancer Res 2004; 64: 1522–1533.

    Article  CAS  Google Scholar 

  17. Gutierrez MC, Detre S, Johnston S, Mohsin SK, Shou J, Allred DC et al. Molecular changes in tamoxifen-resistant breast cancer: relationship between estrogen receptor, HER-2, and p38 mitogen-activated protein kinase. J Clin Oncol 2005; 23: 2469–2476.

    Article  CAS  Google Scholar 

  18. Wang LH, Yang XY, Zhang X, An P, Kim HJ, Huang J et al. Disruption of estrogen receptor DNA-binding domain and related intramolecular communication restores tamoxifen sensitivity in resistant breast cancer. Cancer Cell 2006; 10: 487–499.

    Article  CAS  Google Scholar 

  19. Bayliss J, Hilger A, Vishnu P, Diehl K, El-Ashry D . Reversal of the estrogen receptor negative phenotype in breast cancer and restoration of antiestrogen response. Clin Cancer Res 2007; 13: 7029–7036.

    Article  CAS  Google Scholar 

  20. Hurtado A, Holmes KA, Geistlinger TR, Hutcheson IR, Nicholson RI, Brown M et al. Regulation of ERBB2 by oestrogen receptor-PAX2 determines response to tamoxifen. Nature 2008; 456: 663–666.

    Article  CAS  Google Scholar 

  21. Hurtado A, Holmes KA, Ross-Innes CS, Schmidt D, Carroll JS . FOXA1 is a key determinant of estrogen receptor function and endocrine response. Nat Genet 2011; 43: 27–33.

    Article  CAS  Google Scholar 

  22. Iorns E, Turner NC, Elliott R, Syed N, Garrone O, Gasco M et al. Identification of CDK10 as an important determinant of resistance to endocrine therapy for breast cancer. Cancer Cell 2008; 13: 91–104.

    Article  CAS  Google Scholar 

  23. Musgrove EA, Sutherland RL . Biological determinants of endocrine resistance in breast cancer. Nat Rev Cancer 2009; 9: 631–643.

    Article  CAS  Google Scholar 

  24. Lupien M, Brown M . Cistromics of hormone-dependent cancer. Endocr Relat Cancer 2009; 16: 381–389.

    Article  CAS  Google Scholar 

  25. Hoskins JM, Carey LA, McLeod HL . CYP2D6 and tamoxifen: DNA matters in breast cancer. Nat Rev Cancer 2009; 9: 576–586.

    Article  CAS  Google Scholar 

  26. de Leeuw R, Flach K, Bentin Toaldo C, Alexi X, Canisius S, Neefjes J et al. PKA phosphorylation redirects ERalpha to promoters of a unique gene set to induce tamoxifen resistance. Oncogene 2013; 32: 3543–3551.

    Article  CAS  Google Scholar 

  27. Beelen K, Zwart W, Linn SC . Can predictive biomarkers in breast cancer guide adjuvant endocrine therapy? Nat Rev Clin Oncol 2012; 9: 529–541.

    Article  CAS  Google Scholar 

  28. Bergamaschi A, Christensen BL, Katzenellenbogen BS . Reversal of endocrine resistance in breast cancer: interrelationships among 14-3-3zeta, FOXM1, and a gene signature associated with mitosis. Breast Cancer Res 2011; 13: R70.

    Article  Google Scholar 

  29. Carroll JS, Meyer CA, Song J, Li W, Geistlinger TR, Eeckhoute J et al. Genome-wide analysis of estrogen receptor binding sites. Nat Genet 2006; 38: 1289–1297.

    Article  CAS  Google Scholar 

  30. Welboren WJ, van Driel MA, Janssen-Megens EM, van Heeringen SJ, Sweep FC, Span PN et al. ChIP-Seq of ERalpha and RNA polymerase II defines genes differentially responding to ligands. Embo J 2009; 28: 1418–1428.

    Article  CAS  Google Scholar 

  31. Ross-Innes CS, Stark R, Teschendorff AE, Holmes KA, Ali HR, Dunning MJ et al. Differential oestrogen receptor binding is associated with clinical outcome in breast cancer. Nature 2012; 481: 389–393.

    Article  CAS  Google Scholar 

  32. Cardozo T, Pagano M . The SCF ubiquitin ligase: insights into a molecular machine. Nat Rev Mol Cell Biol 2004; 5: 739–751.

    Article  CAS  Google Scholar 

  33. Lin HK, Wang G, Chen Z, Teruya-Feldstein J, Liu Y, Chan CH et al. Phosphorylation-dependent regulation of cytosolic localization and oncogenic function of Skp2 by Akt/PKB. Nat Cell Biol 2009; 11: 420–432.

    Article  CAS  Google Scholar 

  34. Bhatt S, Xiao Z, Meng Z, Katzenellenbogen BS . Phosphorylation by p38 mitogen-activated protein kinase promotes estrogen receptor alpha turnover and functional activity via the SCF(Skp2) proteasomal complex. Mol Cell Biol 2012; 32: 1928–1943.

    Article  CAS  Google Scholar 

  35. Jin T, Branch DR, Zhang X, Qi S, Youngson B, Goss PE . Examination of POU homeobox gene expression in human breast cancer cells. Int J Cancer 1999; 81: 104–112.

    Article  CAS  Google Scholar 

  36. Pesce M, Scholer HR . Oct-4: gatekeeper in the beginnings of mammalian development. Stem Cells 2001; 19: 271–278.

    Article  CAS  Google Scholar 

  37. Tai MH, Chang CC, Kiupel M, Webster JD, Olson LK, Trosko JE . Oct4 expression in adult human stem cells: evidence in support of the stem cell theory of carcinogenesis. Carcinogenesis 2005; 26: 495–502.

    Article  CAS  Google Scholar 

  38. Loh YH, Wu Q, Chew JL, Vega VB, Zhang W, Chen X et al. The Oct4 and Nanog transcription network regulates pluripotency in mouse embryonic stem cells. Nat Genet 2006; 38: 431–440.

    Article  CAS  Google Scholar 

  39. Monk M, Holding C . Human embryonic genes re-expressed in cancer cells. Oncogene 2001; 20: 8085–8091.

    Article  CAS  Google Scholar 

  40. Gidekel S, Pizov G, Bergman Y, Pikarsky E . Oct-3/4 is a dose-dependent oncogenic fate determinant. Cancer Cell 2003; 4: 361–370.

    Article  CAS  Google Scholar 

  41. Ramos-Mejia V, Escalante-Alcalde D, Kunath T, Ramirez L, Gertsenstein M, Nagy A et al. Phenotypic analyses of mouse embryos with ubiquitous expression of Oct4: effects on mid-hindbrain patterning and gene expression. Dev Dyn 2005; 232: 180–190.

    Article  CAS  Google Scholar 

  42. Nichols J, Zevnik B, Anastassiadis K, Niwa H, Klewe-Nebenius D, Chambers I et al. Formation of pluripotent stem cells in the mammalian embryo depends on the POU transcription factor Oct4. Cell 1998; 95: 379–391.

    Article  CAS  Google Scholar 

  43. Holmes KA, Song JS, Liu XS, Brown M, Carroll JS . Nkx3-1 and LEF-1 function as transcriptional inhibitors of estrogen receptor activity. Cancer Res 2008; 68: 7380–7385.

    Article  CAS  Google Scholar 

  44. Bowen C, Bubendorf L, Voeller HJ, Slack R, Willi N, Sauter G et al. Loss of NKX3.1 expression in human prostate cancers correlates with tumor progression. Cancer Res 2000; 60: 6111–6115.

    CAS  Google Scholar 

  45. Kim MJ, Bhatia-Gaur R, Banach-Petrosky WA, Desai N, Wang Y, Hayward SW et al. Nkx3.1 mutant mice recapitulate early stages of prostate carcinogenesis. Cancer Res 2002; 62: 2999–3004.

    CAS  PubMed  Google Scholar 

  46. Kim MJ, Cardiff RD, Desai N, Banach-Petrosky WA, Parsons R, Shen MM et al. Cooperativity of Nkx3.1 and Pten loss of function in a mouse model of prostate carcinogenesis. Proc Natl Acad Sci USA 2002; 99: 2884–2889.

    Article  CAS  Google Scholar 

  47. Massarweh S, Osborne CK, Creighton CJ, Qin L, Tsimelzon A, Huang S et al. Tamoxifen resistance in breast tumors is driven by growth factor receptor signaling with repression of classic estrogen receptor genomic function. Cancer Res 2008; 68: 826–833.

    Article  CAS  Google Scholar 

  48. Coser KR, Wittner BS, Rosenthal NF, Collins SC, Melas A, Smith SL et al. Antiestrogen-resistant subclones of MCF-7 human breast cancer cells are derived from a common monoclonal drug-resistant progenitor. Proc Natl Acad Sci USA 2009; 106: 14536–14541.

    Article  CAS  Google Scholar 

  49. Allgood VE, Zhang Y, O'Malley BW, Weigel NL . Analysis of chicken progesterone receptor function and phosphorylation using an adenovirus-mediated procedure for high-efficiency DNA transfer. Biochemistry 1997; 36: 224–232.

    Article  CAS  Google Scholar 

  50. Metivier R, Penot G, Hubner MR, Reid G, Brand H, Kos M et al. Estrogen receptor-alpha directs ordered, cyclical, and combinatorial recruitment of cofactors on a natural target promoter. Cell 2003; 115: 751–763.

    Article  CAS  Google Scholar 

  51. Ogawa S, Lozach J, Benner C, Pascual G, Tangirala RK, Westin S et al. Molecular determinants of crosstalk between nuclear receptors and toll-like receptors. Cell 2005; 122: 707–721.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank Dr Peter Yau, Biotechnology Core, UIUC, for contributions to the mass-spectrometry analyses. This work was supported by grants from the NIH 5RO1 CA18119 (BSK), The Breast Cancer Research Foundation (BSK) and Juvenile Diabetes Research Foundation (GW). SB conceived of, designed and executed the study, performed experiments, data analyses and interpretation and wrote the manuscript. JDS conceived the study, performed computational analysis and contributed to experimental data. GW and SJ contributed to experimental data. BSK conceived and designed the study, performed data analyses and interpretation and wrote the manuscript.

Author information

Authors and Affiliations

  1. Departments of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, IL, USA

    S Bhatt & J D Stender

  2. Agilent Technologies, Cedar Creek, TX, USA

    S Joshi

  3. Research Division, Joslin Diabetes Center, Harvard Medical School, Boston, MA, USA

    G Wu

  4. Molecular and Integrative Physiology, University of Illinois at Urbana-Champaign, Urbana, IL, USA

    B S Katzenellenbogen

Authors
  1. S Bhatt
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J D Stender
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. S Joshi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. G Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. B S Katzenellenbogen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to B S Katzenellenbogen.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Additional information

Supplementary Information accompanies this paper on the Oncogene website

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Bhatt, S., Stender, J., Joshi, S. et al. OCT-4: a novel estrogen receptor-α collaborator that promotes tamoxifen resistance in breast cancer cells. Oncogene 35, 5722–5734 (2016). https://doi.org/10.1038/onc.2016.105

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/onc.2016.105

This article is cited by

Search

Advanced search

Quick links