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NFκB signaling is important for growth of antiestrogen resistant breast cancer cells

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Abstract

Resistance to endocrine therapy is a major clinical challenge in current treatment of estrogen receptor-positive breast cancer. The molecular mechanisms underlying resistance are yet not fully clarified. In this study, we investigated whether NFκB signaling is causally involved in antiestrogen resistant cell growth and a potential target for re-sensitizing resistant cells to endocrine therapy. We used an MCF-7-derived cell model for antiestrogen resistant breast cancer to investigate dependence on NFκB signaling for antiestrogen resistant cell growth. We found that targeting NFκB preferentially inhibited resistant cell growth. Antiestrogen resistant cells expressed increased p50 and RelB, and displayed increased phosphorylation of p65 at Ser529 and Ser536. Moreover, transcriptional activity of NFκB after stimulation with tumor necrosis factor α was enhanced in antiestrogen resistant cell lines compared to the parental cell line. Inhibition of NFκB signaling sensitized tamoxifen resistant cells to the growth inhibitory effects of tamoxifen but was not sufficient to fully restore sensitivity of fulvestrant resistant cells to fulvestrant. In support of this, depletion of p65 with siRNA in tamoxifen resistant cells increased sensitivity to tamoxifen treatment. Our data provide evidence that NFκB signaling is enhanced in antiestrogen resistant breast cancer cells and plays an important role for antiestrogen resistant cell growth and for sensitivity to tamoxifen treatment in resistant cells. Our results imply that targeting NFκB might serve as a potential novel treatment strategy for breast cancer patients with resistance toward antiestrogen.

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References

  1. Ali S, Coombes RC (2002) Endocrine-responsive breast cancer and strategies for combating resistance. Nat Rev Cancer 2:101–112

    Article  PubMed  Google Scholar 

  2. Croxtall JD, McKeage K (2011) Fulvestrant: a review of its use in the management of hormone receptor-positive metastatic breast cancer in postmenopausal women. Drugs 71:363–380

    Article  PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  4. Ring A, Dowsett M (2004) Mechanisms of tamoxifen resistance. Endocr Relat Cancer 11:643–658

    Article  PubMed  CAS  Google Scholar 

  5. Frogne T, Benjaminsen RV, Sonne-Hansen K, Sorensen BS, Nexo E, Laenkholm AV, Rasmussen LM, Riese DJ, de Cremoux P, Stenvang J, Lykkesfeldt AE (2009) Activation of ErbB3, EGFR and Erk is essential for growth of human breast cancer cell lines with acquired resistance to fulvestrant. Breast Cancer Res Treat 114:263–275

    Article  PubMed  CAS  Google Scholar 

  6. Frogne T, Laenkholm AV, Lyng MB, Henriksen KL, Lykkesfeldt AE (2009) Determination of HER2 phosphorylation at tyrosine 1221/1222 improves prediction of poor survival for breast cancer patients with hormone receptor-positive tumors. Breast Cancer Res 11:R11

    Article  PubMed  Google Scholar 

  7. Hutcheson IR, Knowlden JM, Madden TA, Barrow D, Gee JM, Wakeling AE, Nicholson RI (2003) Oestrogen receptor-mediated modulation of the EGFR/MAPK pathway in tamoxifen-resistant MCF-7 cells. Breast Cancer Res Treat 81:81–93

    Article  PubMed  CAS  Google Scholar 

  8. Knowlden JM, Hutcheson IR, Jones HE, Madden T, Gee JM, Harper ME, Barrow D, Wakeling AE, Nicholson RI (2003) Elevated levels of epidermal growth factor receptor/c-erbB2 heterodimers mediate an autocrine growth regulatory pathway in tamoxifen-resistant MCF-7 cells. Endocrinology 144:1032–1044

    Article  PubMed  CAS  Google Scholar 

  9. McClelland RA, Barrow D, Madden TA, Dutkowski CM, Pamment J, Knowlden JM, Gee JM, Nicholson RI (2001) Enhanced epidermal growth factor receptor signaling in MCF7 breast cancer cells after long-term culture in the presence of the pure antiestrogen ICI 182,780 (Faslodex). Endocrinology 142:2776–2788

    Article  PubMed  CAS  Google Scholar 

  10. Nicholson RI, Johnston SR (2005) Endocrine therapy—current benefits and limitations. Breast Cancer Res Treat 93(Suppl 1):S3–S10

    Article  PubMed  CAS  Google Scholar 

  11. Gu Z, Lee RY, Skaar TC, Bouker KB, Welch JN, Lu J, Liu A, Zhu Y, Davis N, Leonessa F, Brunner N, Wang Y, Clarke R (2002) Association of interferon regulatory factor-1, nucleophosmin, nuclear factor-kappaB, and cyclic AMP response element binding with acquired resistance to Faslodex (ICI 182,780). Cancer Res 62:3428–3437

    PubMed  CAS  Google Scholar 

  12. Nehra R, Riggins RB, Shajahan AN, Zwart A, Crawford AC, Clarke R (2010) BCL2 and CASP8 regulation by NF-kappaB differentially affect mitochondrial function and cell fate in antiestrogen-sensitive and -resistant breast cancer cells. FASEB J 24:2040–2055

    Article  PubMed  CAS  Google Scholar 

  13. Zhou Y, Yau C, Gray JW, Chew K, Dairkee SH, Moore DH, Eppenberger U, Eppenberger-Castori S, Benz CC (2007) Enhanced NF kappa B and AP-1 transcriptional activity associated with antiestrogen resistant breast cancer. BMC Cancer 7:59

    Article  PubMed  Google Scholar 

  14. deGraffenried LA, Chandrasekar B, Friedrichs WE, Donzis E, Silva J, Hidalgo M, Freeman JW, Weiss GR (2004) NF-kappa B inhibition markedly enhances sensitivity of resistant breast cancer tumor cells to tamoxifen. Ann Oncol 15:885–890

    Article  PubMed  CAS  Google Scholar 

  15. Ghosh S, Karin M (2002) Missing pieces in the NF-kappaB puzzle. Cell 109(Suppl):S81–S96

    Article  PubMed  CAS  Google Scholar 

  16. Hayden MS, Ghosh S (2004) Signaling to NF-kappaB. Genes Dev 18:2195–2224

    Article  PubMed  CAS  Google Scholar 

  17. Viatour P, Merville MP, Bours V, Chariot A (2005) Phosphorylation of NF-kappaB and IkappaB proteins: implications in cancer and inflammation. Trends Biochem Sci 30:43–52

    Article  PubMed  CAS  Google Scholar 

  18. Briand P, Lykkesfeldt AE (1984) Effect of estrogen and antiestrogen on the human breast cancer cell line MCF-7 adapted to growth at low serum concentration. Cancer Res 44:1114–1119

    PubMed  CAS  Google Scholar 

  19. Lykkesfeldt AE, Madsen MW, Briand P (1994) Altered expression of estrogen-regulated genes in a tamoxifen-resistant and ICI 164,384 and ICI 182,780 sensitive human breast cancer cell line, MCF-7/TAMR-1. Cancer Res 54:1587–1595

    PubMed  CAS  Google Scholar 

  20. Lykkesfeldt AE, Larsen SS, Briand P (1995) Human breast cancer cell lines resistant to pure anti-estrogens are sensitive to tamoxifen treatment. Int J Cancer 61:529–534

    Article  PubMed  CAS  Google Scholar 

  21. Lundholt BK, Briand P, Lykkesfeldt AE (2001) Growth inhibition and growth stimulation by estradiol of estrogen receptor transfected human breast epithelial cell lines involve different pathways. Breast Cancer Res Treat 67:199–214

    Article  PubMed  CAS  Google Scholar 

  22. Sonne-Hansen K, Norrie IC, Emdal KB, Benjaminsen RV, Frogne T, Christiansen IJ, Kirkegaard T, Lykkesfeldt AE (2010) Breast cancer cells can switch between estrogen receptor alpha and ErbB signaling and combined treatment against both signaling pathways postpones development of resistance. Breast Cancer Res Treat 121:601–613

    Article  PubMed  CAS  Google Scholar 

  23. Dietrich N, Thastrup J, Holmberg C, Gyrd-Hansen M, Fehrenbacher N, Lademann U, Lerdrup M, Herdegen T, Jaattela M, Kallunki T (2004) JNK2 mediates TNF-induced cell death in mouse embryonic fibroblasts via regulation of both caspase and cathepsin protease pathways. Cell Death Differ 11:301–313

    Article  PubMed  CAS  Google Scholar 

  24. Yde CW, Olsen BB, Meek D, Watanabe N, Guerra B (2008) The regulatory beta-subunit of protein kinase CK2 regulates cell-cycle progression at the onset of mitosis. Oncogene 27:4986–4997

    Article  PubMed  CAS  Google Scholar 

  25. Waelchli R, Bollbuck B, Bruns C, Buhl T, Eder J, Feifel R, Hersperger R, Janser P, Revesz L, Zerwes HG, Schlapbach A (2006) Design and preparation of 2-benzamido-pyrimidines as inhibitors of IKK. Bioorg Med Chem Lett 16:108–112

    Article  PubMed  CAS  Google Scholar 

  26. Zhou Y, Eppenberger-Castori S, Eppenberger U, Benz CC (2005) The NFkappaB pathway and endocrine-resistant breast cancer. Endocr Relat Cancer 12(Suppl 1):S37–S46

    Article  PubMed  CAS  Google Scholar 

  27. Shen HM, Tergaonkar V (2009) NFkappaB signaling in carcinogenesis and as a potential molecular target for cancer therapy. Apoptosis 14:348–363

    Article  PubMed  CAS  Google Scholar 

  28. Madrid LV, Mayo MW, Reuther JY, Baldwin AS Jr (2001) Akt stimulates the transactivation potential of the RelA/p65 Subunit of NF-kappa B through utilization of the Ikappa B kinase and activation of the mitogen-activated protein kinase p38. J Biol Chem 276:18934–18940

    Article  PubMed  CAS  Google Scholar 

  29. Frogne T, Jepsen JS, Larsen SS, Fog CK, Brockdorff BL, Lykkesfeldt AE (2005) Antiestrogen-resistant human breast cancer cells require activated protein kinase B/Akt for growth. Endocr Relat Cancer 12:599–614

    Article  PubMed  CAS  Google Scholar 

  30. Pradhan M, Bembinster LA, Baumgarten SC, Frasor J (2010) Proinflammatory cytokines enhance estrogen-dependent expression of the multidrug transporter gene ABCG2 through estrogen receptor and NF{kappa}B cooperativity at adjacent response elements. J Biol Chem 285:31100–31106

    Article  PubMed  CAS  Google Scholar 

  31. Nettles KW, Gil G, Nowak J, Métivier R, Sharma VB, Greene GL (2008) CBP Is a dosage-dependent regulator of nuclear factor-kappaB suppression by the estrogen receptor. Mol Endocrinol 22:263–272

    Article  PubMed  CAS  Google Scholar 

  32. Madsen MW, Reiter BE, Lykkesfeldt AE (1995) Differential expression of estrogen receptor mRNA splice variants in the tamoxifen resistant human breast cancer cell line, MCF-7/TAMR-1 compared to the parental MCF-7 cell line. Mol Cell Endocrinol 109:197–207

    Article  PubMed  CAS  Google Scholar 

  33. Larsen SS, Madsen MW, Jensen BL, Lykkesfeldt AE (1997) Resistance of human breast-cancer cells to the pure steroidal anti-estrogen ICI 182,780 is not associated with a general loss of estrogen-receptor expression or lack of estrogen responsiveness. Int J Cancer 72:1129–1136

    Article  PubMed  CAS  Google Scholar 

  34. Cutrupi S, Reineri S, Panetto A, Grosso E, Caizzi L, Ricci L, Friard O, Agati S, Scatolini M, Chiorino G, Lykkesfeldt AE, De Bortoli M (2012) Targeting of the adaptor protein Tab 2 as a novel approach to revert tamoxifen resistance in breast cancer cells. Oncogene. doi:10.1038/onc.2011.627

  35. Takaesu G, Kishida S, Hiyama A, Yamaguchi K, Shibuya H, Irie K, Ninomiya-Tsuji J, Matsumoto K (2000) TAB 2, a novel adaptor protein, mediates activation of TAK1 MAPKKK by linking TAK1 to TRAF6 in the IL-1 signal transduction pathway. Mol Cell 5:649–658

    Article  PubMed  CAS  Google Scholar 

  36. Riggins RB, Zwart A, Nehra R, Clarke R (2005) The nuclear factor kappa B inhibitor parthenolide restores ICI 182,780 (Faslodex; fulvestrant)-induced apoptosis in antiestrogen-resistant breast cancer cells. Mol Cancer Ther 4:33–41

    PubMed  CAS  Google Scholar 

  37. Buss H, Dorrie A, Schmitz ML, Hoffmann E, Resch K, Kracht M (2004) Constitutive and interleukin-1-inducible phosphorylation of p65 NF-{kappa}B at serine 536 is mediated by multiple protein kinases including I{kappa}B kinase (IKK)-{alpha}, IKK{beta}, IKK{epsilon}, TRAF family member-associated (TANK)-binding kinase 1 (TBK1), and an unknown kinase and couples p65 to TATA-binding protein-associated factor II31-mediated interleukin-8 transcription. J Biol Chem 279:55633–55643

    Article  PubMed  CAS  Google Scholar 

  38. Song YJ, Jen KY, Soni V, Kieff E, Cahir-McFarland E (2006) IL-1 receptor-associated kinase 1 is critical for latent membrane protein 1-induced p65/RelA serine 536 phosphorylation and NF-kappaB activation. Proc Natl Acad Sci USA 103:2689–2694

    Article  PubMed  CAS  Google Scholar 

  39. Wang D, Westerheide SD, Hanson JL, Baldwin AS Jr (2000) Tumor necrosis factor alpha-induced phosphorylation of RelA/p65 on Ser529 is controlled by casein kinase II. J Biol Chem 275:32592–32597

    Article  PubMed  CAS  Google Scholar 

  40. McElhinny JA, Trushin SA, Bren GD, Chester N, Paya CV (1996) Casein kinase II phosphorylates I kappa B alpha at S-283, S-289, S-293, and T-291 and is required for its degradation. Mol Cell Biol 16:899–906

    PubMed  CAS  Google Scholar 

  41. Yde CW, Frogne T, Lykkesfeldt AE, Fichtner I, Issinger OG, Stenvang J (2007) Induction of cell death in antiestrogen resistant human breast cancer cells by the protein kinase CK2 inhibitor DMAT. Cancer Lett 256:229–237

    Article  PubMed  CAS  Google Scholar 

  42. Wang X, Belguise K, Kersual N, Kirsch KH, Mineva ND, Galtier F, Chalbos D, Sonenshein GE (2007) Oestrogen signalling inhibits invasive phenotype by repressing RelB and its target BCL2. Nat Cell Biol 9:470–478

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

We thank Marja Jäättelä for kindly providing pNFκB-Luc and pRL-null plasmids. We thank Jane Lind Christensen, Birgit Reiter, and Inger Heiberg for excellent technical assistance. This study was supported by a grant from the Danish Medical Research Council (FSS 271-07-0704) to CWY.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Breast Cancer Group, Cell Death and Metabolism, Danish Cancer Society Research Center, Strandboulevarden 49, 2100, Copenhagen Ø, Denmark

    Christina W. Yde, Kristina B. Emdal & Anne E. Lykkesfeldt

  2. Department of Biochemistry and Molecular Biology, University of Southern Denmark, Campusvej 55, 5230, Odense M, Denmark

    Barbara Guerra

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  1. Christina W. Yde
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  2. Kristina B. Emdal
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  3. Barbara Guerra
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  4. Anne E. Lykkesfeldt
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Correspondence to Christina W. Yde.

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Yde, C.W., Emdal, K.B., Guerra, B. et al. NFκB signaling is important for growth of antiestrogen resistant breast cancer cells. Breast Cancer Res Treat 135, 67–78 (2012). https://doi.org/10.1007/s10549-012-2053-1

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  • DOI: https://doi.org/10.1007/s10549-012-2053-1

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