Article | Published:

c-Jun/AP-1 overexpression reprograms ERα signaling related to tamoxifen response in ERα-positive breast cancer

Oncogenevolume 37pages25862600 (2018) | Download Citation


A critical mechanism that has been proposed for transcription regulation by estrogen receptor α (ER) is the tethering of ER to DNA via other transcription factors, such as AP-1. However, genome-wide assessment of the overlap in chromatin binding repertoires of these two transcription factors has not been reported. Here, we show that the AP-1 transcription factor c-Jun interacts with ER and that c-Jun chromatin binding shows extensive overlap with ER binding at the global level. Further, we show that c-Jun overexpression reprograms ER chromatin binding and modulates ER-mediated gene regulation. Our data are consistent with a mechanism where estrogen/ER-dependent crosstalk with AP-1 at the transcriptional level is mediated through the tethering of ER to DNA bound AP-1. Additionally, in our system c-Jun overexpression causes reduced sensitivity to tamoxifen in ER+ breast cancer cells. Integrated cistrome, transcriptome, and clinical data reveal TGFBI as a candidate gene which may confer tamoxifen resistance by ER and AP-1 crosstalk. Further, we show that TGFBI expression is elevated in breast cancer compared to normal breast. Together, our data provide a novel genome-wide footprint of ER and AP-1 crosstalk and suggest AP-1 and TGFBI signaling as potential therapeutic targets in AP-1-overexpressing ER-positive breast tumors.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.


  1. 1.

    Zhao C, Dahlman-Wright K, Gustafsson JA. Estrogen signaling via estrogen receptor β. J Biol Chem. 2010;285:39575–9.

  2. 2.

    den Hollander P, Savage MI, Brown PH. Targeted therapy for breast cancer prevention. Front Oncol. 2013;3:250.

  3. 3.

    Davies C, Godwin J, Gray R, Clarke M, Cutter D, Darby S, et al. Relevance of breast cancer hormone receptors and other factors to the efficacy of adjuvant tamoxifen: patient-level meta-analysis of randomised trials. Lancet. 2011;378:771–84.

  4. 4.

    Musgrove EA, Sutherland RL. Biological determinants of endocrine resistance in breast cancer. Nat Rev Cancer. 2009;9:631–43.

  5. 5.

    Osborne CK, Schiff R. Mechanisms of endocrine resistance in breast cancer. Annu Rev Med. 2011;62:233–47.

  6. 6.

    Ali S, Rasool M, Chaoudhry H, P NP, Jha P, Hafiz A, et al. Molecular mechanisms and mode of tamoxifen resistance in breast cancer. Bioinformation. 2016;12:135–9.

  7. 7.

    Malorni L, Giuliano M, Migliaccio I, Wang T, Creighton CJ, Lupien M, et al. Blockade of AP-1 potentiates endocrine therapy and overcomes resistance. Mol Cancer Res. 2016;14:470–81.

  8. 8.

    Shaulian E. AP-1—The Jun proteins: oncogenes or tumor suppressors in disguise? Cell Signal. 2010;22:894–9.

  9. 9.

    Johnston SR, Lu B, Scott GK, Kushner PJ, Smith IE, Dowsett M, et al. Increased activator protein-1 DNA binding and c-Jun NH2-terminal kinase activity in human breast tumors with acquired tamoxifen resistance. Clin Cancer Res. 1999;5:251–6.

  10. 10.

    Cheung E, Acevedo ML, Cole PA, Kraus WL. Altered pharmacology and distinct coactivator usage for estrogen receptor-dependent transcription through activating protein-1. Proc Natl Acad Sci USA. 2005;102:559–64.

  11. 11.

    Safe S. Transcriptional activation of genes by 17 beta-estradiol through estrogen receptor-Sp1 interactions. Vitam Horm. 2001;62:231–52.

  12. 12.

    Luo M, Koh M, Feng J, Wu Q, Melamed P. Cross talk in hormonally regulated gene transcription through induction of estrogen receptor ubiquitylation. Mol Cell Biol. 2005;25:7386–98.

  13. 13.

    Stender JD, Kim K, Charn TH, Komm B, Chang KC, Kraus WL, et al. Genome-wide analysis of estrogen receptor alpha DNA binding and tethering mechanisms identifies Runx1 as a novel tethering factor in receptor-mediated transcriptional activation. Mol Cell Biol. 2010;30:3943–55.

  14. 14.

    Heldring N, Isaacs GD, Diehl AG, Sun M, Cheung E, Ranish JA, et al. Multiple sequence-specific DNA-binding proteins mediate estrogen receptor signaling through a tethering pathway. Mol Endocrinol. 2011;25:564–74.

  15. 15.

    Lupien M, Meyer CA, Bailey ST, Eeckhoute J, Cook J, Westerling T, et al. Growth factor stimulation induces a distinct ER(alpha) cistrome underlying breast cancer endocrine resistance. Genes Dev. 2010;24:2219–27.

  16. 16.

    Tiniakos DG, Scott LE, Corbett IP, Piggott NH, Horne CH. Studies of c-jun oncogene expression in human breast using a new monoclonal antibody, NCL-DK4. J Pathol. 1994;172:19–26.

  17. 17.

    Zhao C, Qiao Y, Jonsson P, Wang J, Xu L, Rouhi P, et al. Genome-wide profiling of AP-1-regulated transcription provides insights into the invasiveness of triple-negative breast cancer. Cancer Res. 2014;74:3983–94.

  18. 18.

    Eferl R, Wagner EF. AP-1: a double-edged sword in tumorigenesis. Nat Rev Cancer. 2003;3:859–68.

  19. 19.

    Zhu J, Zhao C, Kharman-Biz A, Zhuang T, Jonsson P, Liang N, et al. The atypical ubiquitin ligase RNF31 stabilizes estrogen receptor alpha and modulates estrogen-stimulated breast cancer cell proliferation. Oncogene. 2014;33:4340–51.

  20. 20.

    Zhang Y, Pu X, Shi M, Chen L, Song Y, Qian L, et al. Critical role of c-Jun overexpression in liver metastasis of human breast cancer xenograft model. BMC Cancer. 2007;7:145.

  21. 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.

  22. 22.

    Ip YT, Davis RJ. Signal transduction by the c-Jun N-terminal kinase (JNK)—from inflammation to development. Curr Opin Cell Biol. 1998;10:205–19.

  23. 23.

    Sun M, Isaacs GD, Hah N, Heldring N, Fogarty EA, Kraus WL. Estrogen regulates JNK1 genomic localization to control gene expression and cell growth in breast cancer cells. Mol Endocrinol. 2012;26:736–47.

  24. 24.

    Lupien M, Eeckhoute J, Meyer CA, Wang Q, Zhang Y, Li W, et al. FoxA1 translates epigenetic signatures into enhancer-driven lineage-specific transcription. Cell. 2008;132:958–70.

  25. 25.

    Kong SL, Li G, Loh SL, Sung WK, Liu ET. Cellular reprogramming by the conjoint action of ERalpha, FOXA1, and GATA3 to a ligand-inducible growth state. Mol Syst Biol. 2011;7:526.

  26. 26.

    Jia M, Andreassen T, Jensen L, Bathen TF, Sinha I, Gao H, et al. Estrogen receptor alpha promotes breast cancer by reprogramming choline metabolism. Cancer Res. 2016;76:5634–46.

  27. 27.

    Gyorffy B, Lanczky A, Eklund AC, Denkert C, Budczies J, Li Q, et al. An online survival analysis tool to rapidly assess the effect of 22,277 genes on breast cancer prognosis using microarray data of 1,809 patients. Breast Cancer Res Treat. 2010;123:725–31.

  28. 28.

    Curtis C, Shah SP, Chin SF, Turashvili G, Rueda OM, Dunning MJ, et al. The genomic and transcriptomic architecture of 2000 breast tumours reveals novel subgroups. Nature. 2012;486:346–52.

  29. 29.

    Ma C, Rong Y, Radiloff DR, Datto MB, Centeno B, Bao S, et al. Extracellular matrix protein betaig-h3/TGFBI promotes metastasis of colon cancer by enhancing cell extravasation. Genes Dev. 2008;22:308–21.

  30. 30.

    Schneider D, Kleeff J, Berberat PO, Zhu Z, Korc M, Friess H, et al. Induction and expression of betaig-h3 in pancreatic cancer cells. Biochim Biophys Acta. 2002;1588:1–6.

  31. 31.

    Shen Q, Bai Y, Chang KC, Wang Y, Burris TP, Freedman LP, et al. Liver X receptor-retinoid X receptor (LXR-RXR) heterodimer cistrome reveals coordination of LXR and AP1 signaling in keratinocytes. J Biol Chem. 2011;286:14554–63.

  32. 32.

    Biddie SC, John S, Sabo PJ, Thurman RE, Johnson TA, Schiltz RL, et al. Transcription factor AP1 potentiates chromatin accessibility and glucocorticoid receptor binding. Mol Cell. 2011;43:145–55.

  33. 33.

    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;481:389–93.

  34. 34.

    Ji Z, Donaldson IJ, Liu J, Hayes A, Zeef LA, Sharrocks AD. The forkhead transcription factor FOXK2 promotes AP-1-mediated transcriptional regulation. Mol Cell Biol. 2012;32:385–98.

  35. 35.

    Theodorou V, Stark R, Menon S, Carroll JS. GATA3 acts upstream of FOXA1 in mediating ESR1 binding by shaping enhancer accessibility. Genome Res. 2013;23:12–22.

  36. 36.

    Jin HJ, Zhao JC, Wu L, Kim J, Yu J. Cooperativity and equilibrium with FOXA1 define the androgen receptor transcriptional program. Nat Commun. 2014;5:3972.

  37. 37.

    Robinson JL, Hickey TE, Warren AY, Vowler SL, Carroll T, Lamb AD, et al. Elevated levels of FOXA1 facilitate androgen receptor chromatin binding resulting in a CRPC-like phenotype. Oncogene. 2014;33:5666–74.

  38. 38.

    Wang D, Garcia-Bassets I, Benner C, Li W, Su X, Zhou Y, et al. Reprogramming transcription by distinct classes of enhancers functionally defined by eRNA. Nature. 2011;474:390–4.

  39. 39.

    Fu X, Jeselsohn R, Pereira R, Hollingsworth EF, Creighton CJ, Li F, et al. FOXA1 overexpression mediates endocrine resistance by altering the ER transcriptome and IL-8 expression in ER-positive breast cancer. Proc Natl Acad Sci USA. 2016;113:E6600–9.

  40. 40.

    Stender JD, Nwachukwu JC, Kastrati I, Kim Y, Strid T, Yakir M, et al. Structural and Molecular Mechanisms of Cytokine-Mediated Endocrine Resistance in Human Breast Cancer Cells. Mol Cell. 2017;65:1122–35. e1125

  41. 41.

    Wright TM, Wardell SE, Jasper JS, Stice JP, Safi R, Nelson ER, et al. Delineation of a FOXA1/ERalpha/AGR2 regulatory loop that is dysregulated in endocrine therapy-resistant breast cancer. Mol Cancer Res. 2014;12:1829–39.

  42. 42.

    Brown HA, Thomas PG, Lindsley CW. Targeting phospholipase D in cancer, infection and neurodegenerative disorders. Nat Rev Drug Discov. 2017;16:351–67.

  43. 43.

    Chen Y, Zheng Y, Foster DA. Phospholipase D confers rapamycin resistance in human breast cancer cells. Oncogene. 2003;22:3937–42.

  44. 44.

    Kharman-Biz A, Gao H, Ghiasvand R, Zhao C, Zendehdel K, Dahlman-Wright K. Expression of activator protein-1 (AP-1) family members in breast cancer. BMC Cancer. 2013;13:441.

  45. 45.

    Ween MP, Oehler MK, Ricciardelli C. Transforming growth Factor-Beta-Induced Protein (TGFBI)/(betaig-H3): a matrix protein with dual functions in ovarian cancer. Int J Mol Sci. 2012;13:10461–77.

  46. 46.

    Ahmed AA, Mills AD, Ibrahim AE, Temple J, Blenkiron C, Vias M, et al. The extracellular matrix protein TGFBI induces microtubule stabilization and sensitizes ovarian cancers to paclitaxel. Cancer Cell. 2007;12:514–27.

  47. 47.

    Brunen D, Willems SM, Kellner U, Midgley R, Simon I, Bernards R. TGF-beta: an emerging player in drug resistance. Cell Cycle (Georget, Tex. 2013;12:2960–8.

  48. 48.

    Neuzillet C, Tijeras-Raballand A, Cohen R, Cros J, Faivre S, Raymond E, et al. Targeting the TGFbeta pathway for cancer therapy. Pharmacol Ther. 2015;147:22–31.

  49. 49.

    Zhao C, Matthews J, Tujague M, Wan J, Strom A, Toresson G, et al. Estrogen receptor beta2 negatively regulates the transactivation of estrogen receptor alpha in human breast cancer cells. Cancer Res. 2007;67:3955–62.

  50. 50.

    Heinz S, Benner C, Spann N, Bertolino E, Lin YC, Laslo P, et al. Simple combinations of lineage-determining transcription factors prime cis-regulatory elements required for macrophage and B cell identities. Mol Cell. 2010;38:576–89.

  51. 51.

    Dahlman-Wright K, Qiao Y, Jonsson P, Gustafsson JA, Williams C, Zhao C. Interplay between AP-1 and estrogen receptor alpha in regulating gene expression and proliferation networks in breast cancer cells. Carcinogenesis. 2012;33:1684–91.

  52. 52.

    Hung MS, Chen IC, You L, Jablons DM, Li YC, Mao JH, et al. Knockdown of Cul4A increases chemosensitivity to gemcitabine through upregulation of TGFBI in lung cancer cells. Oncol Rep. 2015;34:3187–95.

Download references


The authors thank the Bioinformatic and Expression Analysis core facility at the Karolinska Institute ( for performing the Affymetrix and ChIP-seq assays and Hui Gao for critically reading the manuscript. This work was supported by scholarship from the China Scholarship Council, a PhD student grant (KID) from the Karolinska Institutet and grants from the Swedish Cancer Society (Cancerfonden).

Author contributions

HH and CZ performed experiments; RF prepared materials; IS and CZ analyzed data; HH, CZ, and KD interpreted results of experiments; HH, CZ, FY, and KD prepared figures; HH and CZ drafted manuscript; HH, L-AH, CZ, and KD edited and revised manuscript; all authors approved final version of manuscript; CZ and KD initiated and designed the study and supervised HH.

Author information


  1. Department of Biosciences and Nutrition, Novum, Karolinska Institute, Huddinge, SE-141 83, Sweden

    • Huan He
    • , Indranil Sinha
    • , Rongrong Fan
    • , Lars-Arne Haldosen
    • , Feifei Yan
    • , Chunyan Zhao
    •  & Karin Dahlman-Wright


  1. Search for Huan He in:

  2. Search for Indranil Sinha in:

  3. Search for Rongrong Fan in:

  4. Search for Lars-Arne Haldosen in:

  5. Search for Feifei Yan in:

  6. Search for Chunyan Zhao in:

  7. Search for Karin Dahlman-Wright in:

Conflict of interest

The authors declare that they have no conflict of interest.

Corresponding author

Correspondence to Chunyan Zhao.

Electronic supplementary material

About this article

Publication history





Issue Date