Abstract
ErbB-2 overexpression in breast tumors is associated with poor survival. Expression of Notch-1 and its ligand, Jagged-1, is associated with the poorest survival, including ErbB-2-positive tumors. Trastuzumab plus chemotherapy is the standard of care for ErbB-2-positive breast cancer. A proportion of tumors are initially resistant to trastuzumab and acquired resistance to trastuzumab occurs in metastatic breast cancer and is associated with poor prognosis. Thus, we investigated whether Notch-1 contributes to trastuzumab resistance. ErbB-2-positive cells have low Notch transcriptional activity compared to non-overexpressing cells. Trastuzumab or a dual epidermal growth factor receptor (EGFR)/ErbB-2 tyrosine kinase inhibitor (TKI) increased Notch activity by 2- to 6-fold in SKBr3, BT474 and MCF-7/HER2-18 cells. The increase in activity was abrogated by a Notch inhibitor, γ-secretase inhibitor (GSI) or Notch-1 small-interfering RNA (siRNA). Trastuzumab decreased Notch-1™ precursor, increased amount and nuclear accumulation of active Notch-1IC and increased expression of targets, Hey1 and Deltex1 mRNAs, and Hes5, Hey1, Hes1 proteins. Importantly, trastuzumab-resistant BT474 cells treated with trastuzumab for 6 months expressed twofold higher Notch-1, twofold higher Hey1, ninefold higher Deltex1 mRNAs and threefold higher Notch-1 and Hes5 proteins, compared to trastuzumab-sensitive BT474 cells. The increase in Hey1 and Deltex1 mRNAs in resistant cells was abrogated by a Notch-1 siRNA. Cell proliferation was inhibited more effectively by trastuzumab or TKI plus a GSI than either agent alone. Decreased Notch-1 by siRNA increased efficacy of trastuzumab in BT474 sensitive cells and restored sensitivity in resistant cells. Trastuzumab plus a GSI increased apoptosis in sensitive cells by 20–30%. A GSI alone was sufficient to increase apoptosis in trastuzumab-resistant BT474 cells by 20%, which increased to 30% with trastuzumab. Notch-1 siRNA alone decreased cell growth by 30% in sensitive and more than 50% in resistant BT474 cells. Furthermore, growth of both trastuzumab sensitive and resistant cells was completely inhibited by combining trastuzumab plus Notch-1 siRNA. More importantly, Notch-1 siRNA or a GSI resensitized trastuzumab-resistant BT474 cells to trastuzumab. These results demonstrate that ErbB-2 overexpression suppresses Notch-1 activity, which can be reversed by trastuzumab or TKI. These results suggest that Notch-1 might play a novel role in resistance to trastuzumab, which could be prevented or reversed by inhibiting Notch-1.
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References
Amar S, Moreno-Aspitia A, Perez EA . (2008). Issues and controversies in the treatment of HER2 positive metastatic breast cancer. Breast Cancer Res Treat 109: 1–7.
Arpino G, Weiss H, Lee AV, Schiff R, De Placido S, Osborne CK et al. (2005). Estrogen receptor-positive, progesterone receptor-negative breast cancer: association with growth factor receptor expression and tamoxifen resistance. J Natl Cancer Inst 97: 1254–1261.
Artavanis-Tsakonas S, Rand MD, Lake RJ . (1999). Notch signaling: cell fate control and signal integration in development. Science 284: 770–776.
Baonza A, Freeman M . (2005). Control of cell proliferation in the Drosophila eye by Notch signaling. Dev Cell 8: 529–539.
Baselga J, Albanell J, Molina MA, Arribas J . (2001). Mechanism of action of trastuzumab and scientific update. Semin Oncol 28: 4–11.
Benz CC, Scott GK, Sarup JC, Johnson RM, Tripathy D, Coronado E et al. (1992). Estrogen-dependent, tamoxifen-resistant tumorigenic growth of MCF-7 cells transfected with HER2/neu. Breast Cancer Res Treat 24: 85–95.
Blaumueller CM, Qi H, Zagouras P, Artavanis-Tsakonas S . (1997). Intracellular cleavage of Notch leads to a heterodimeric receptor on the plasma membrane. Cell 90: 281–291.
Brou C, Logeat F, Gupta N, Bessia C, LeBail O, Doedens JR et al. (2000). A novel proteolytic cleavage involved in Notch signaling: the role of the disintegrin-metalloprotease TACE. Mol Cell 5: 207–216.
Burstein HJ, Harris LN, Marcom PK, Lambert-Falls R, Havlin K, Overmoyer B et al. (2003). Trastuzumab and vinorelbine as first-line therapy for HER2-overexpressing metastatic breast cancer: multicenter phase II trial with clinical outcomes, analysis of serum tumor markers as predictive factors, and cardiac surveillance algorithm. J Clin Oncol 21: 2889–2895.
Burstein HJ, Kuter I, Campos SM, Gelman RS, Tribou L, Parker LM et al. (2001). Clinical activity of trastuzumab and vinorelbine in women with HER2-overexpressing metastatic breast cancer. J Clin Oncol 19: 2722–2730.
Calzavara E, Chiaramonte R, Cesana D, Basile A, Sherbet GV, Comi P . (2007). Reciprocal regulation of Notch and PI3K/Akt signalling in T-ALL cells in vitro. J Cell Biochem 103: 1405–1412.
Carter P, Presta L, Gorman CM, Ridgway JB, Henner D, Wong WL et al. (1992). Humanization of an anti-p185HER2 antibody for human cancer therapy. Proc Natl Acad Sci USA 89: 4285–4289.
Cobleigh MA, Vogel CL, Tripathy D, Robert NJ, Scholl S, Fehrenbacher L et al. (1999). Multinational study of the efficacy and safety of humanized anti-HER2 monoclonal antibody in women who have HER2-overexpressing metastatic breast cancer that has progressed after chemotherapy for metastatic disease. J Clin Oncol 17: 2639–2648.
Dickson BC, Mulligan AM, Zhang H, Lockwood G, O’Malley FP, Egan SE et al. (2007). High-level JAG1 mRNA and protein predict poor outcome in breast cancer. Mod Pathol 20: 685–693.
Dievart A, Beaulieu N, Jolicoeur P . (1999). Involvement of Notch1 in the development of mouse mammary tumors. Oncogene 18: 5973–5981.
Dontu G, Jackson KW, McNicholas E, Kawamura MJ, Abdallah WM, Wicha MS . (2004). Role of Notch signaling in cell-fate determination of human mammary stem/progenitor cells. Breast Cancer Res 6: R605–R615.
Efstratiadis A, Szabolcs M, Klinakis A . (2007). Notch, Myc and breast cancer. Cell Cycle 6: 418–429.
Farnie G, Clarke RB, Spence K, Pinnock N, Brennan K, Anderson NG et al. (2007). Novel cell culture technique for primary ductal carcinoma in situ: role of Notch and epidermal growth factor receptor signaling pathways. J Natl Cancer Inst 99: 616–627.
Florena AM, Tripodo C, Guarnotta C, Ingrao S, Porcasi R, Martorana A et al. (2007). Associations between Notch-2, Akt-1 and HER2/neu expression in invasive human breast cancer: a tissue microarray immunophenotypic analysis on 98 patients. Pathobiology 74: 317–322.
Geyer CE, Forster J, Lindquist D, Chan S, Romieu CG, Pienkowski T et al. (2006). Lapatinib plus capecitabine for HER2-positive advanced breast cancer. N Engl J Med 355: 2733–2743.
Imatani A, Callahan R . (2000). Identification of a novel NOTCH-4/INT-3 RNA species encoding an activated gene product in certain human tumor cell lines. Oncogene 19: 223–231.
Jahanzeb M, Mortimer JE, Yunus F, Irwin DH, Speyer J, Koletsky AJ et al. (2002). Phase II trial of weekly vinorelbine and trastuzumab as first-line therapy in patients with HER2(+) metastatic breast cancer. Oncologist 7: 410–417.
Lindsell CE, Shawber CJ, Boulter J, Weinmaster G . (1995). Jagged: a mammalian ligand that activates Notch1. Cell 80: 909–917.
Logeat F, Bessia C, Brou C, LeBail O, Jarriault S, Seidah NG et al. (1998). The Notch1 receptor is cleaved constitutively by a furin-like convertase. Proc Natl Acad Sci USA 95: 8108–8112.
Lu Y, Zi X, Pollak M . (2004). Molecular mechanisms underlying IGF-I-induced attenuation of the growth-inhibitory activity of trastuzumab (Herceptin) on SKBR3 breast cancer cells. Int J Cancer 108: 334–341.
Menendez JA, Mehmi I, Lupu R . (2006). Trastuzumab in combination with heregulin-activated Her-2 (erbB-2) triggers a receptor-enhanced chemosensitivity effect in the absence of Her-2 overexpression. J Clin Oncol 24: 3735–3746.
Miele L . (2006). Notch signaling. Clin Cancer Res 12: 1074–1079.
Miele L, Golde T, Osborne B . (2006). Notch signaling in cancer. Curr Mol Med 6: 905–918.
Miele L, Osborne B . (1999). Arbiter of differentiation and death: Notch signaling meets apoptosis. J Cell Physiol 181: 393–409.
Mumm JS, Schroeter EH, Saxena MT, Griesemer A, Tian X, Pan DJ et al. (2000). A ligand-induced extracellular cleavage regulates gamma-secretase-like proteolytic activation of Notch1. Mol Cell 5: 197–206.
Muss HB, Thor AD, Berry DA, Kute T, Liu ET, Koerner F et al. (1994). c-erbB-2 expression and response to adjuvant therapy in women with node-positive early breast cancer. N Engl J Med 330: 1260–1266.
Nabholtz JM, Slamon D . (2001). New adjuvant strategies for breast cancer: meeting the challenge of integrating chemotherapy and trastuzumab (Herceptin). Semin Oncol 28: 1–12.
Nagata Y, Lan KH, Zhou X, Tan M, Esteva FJ, Sahin AA et al. (2004). PTEN activation contributes to tumor inhibition by trastuzumab, and loss of PTEN predicts trastuzumab resistance in patients. Cancer Cell 6: 117–127.
Nahta R, Esteva FJ . (2006). Herceptin: mechanisms of action and resistance. Cancer Lett 232: 123–138.
Nahta R, Yuan LX, Zhang B, Kobayashi R, Esteva FJ . (2005). Insulin-like growth factor-I receptor/human epidermal growth factor receptor 2 heterodimerization contributes to trastuzumab resistance of breast cancer cells. Cancer Res 65: 11118–11128.
O’Neill CF, Urs S, Cinelli C, Lincoln A, Nadeau RJ, Leon R et al. (2007). Notch2 signaling induces apoptosis and inhibits human MDA-MB-231 xenograft growth. Am J Pathol 171: 1023–1036.
Osborne CK, Bardou V, Hopp TA, Chamness GC, Hilsenbeck SG, Fuqua SA et al. (2003). Role of the estrogen receptor coactivator AIB1 (SRC-3) and HER-2/neu in tamoxifen resistance in breast cancer. J Natl Cancer Inst 95: 353–361.
Osipo C, Gajdos C, Liu H, Chen B, Jordan VC . (2003). Paradoxical action of fulvestrant in estradiol-induced regression of tamoxifen-stimulated breast cancer. J Natl Cancer Inst 95: 1597–1608.
Oswald F, Liptay S, Adler G, Schmid RM . (1998). NF-kappaB2 is a putative target gene of activated Notch-1 via RBP-Jkappa. Mol Cell Biol 18: 2077–2088.
Papaldo P, Fabi A, Ferretti G, Mottolese M, Cianciulli AM, Di Cocco B et al. (2006). A phase II study on metastatic breast cancer patients treated with weekly vinorelbine with or without trastuzumab according to HER2 expression: changing the natural history of HER2-positive disease. Ann Oncol 17: 630–636.
Parks AL, Klueg KM, Stout JR, Muskavitch MA . (2000). Ligand endocytosis drives receptor dissociation and activation in the Notch pathway. Development 127: 1373–1385.
Phillips TM, McBride WH, Pajonk F . (2006). The response of CD24(−/low)/CD44+ breast cancer-initiating cells to radiation. J Natl Cancer Inst 98: 1777–1785.
Politi K, Feirt N, Kitajewski J . (2004). Notch in mammary gland development and breast cancer. Semin Cancer Biol 14: 341–347.
Price-Schiavi SA, Jepson S, Li P, Arango M, Rudland PS, Yee L et al. (2002). Rat Muc4 (sialomucin complex) reduces binding of anti-ErbB2 antibodies to tumor cell surfaces, a potential mechanism for herceptin resistance. Int J Cancer 99: 783–791.
Rangarajan A, Syal R, Selvarajah S, Chakrabarti O, Sarin A, Krishna S . (2001). Activated Notch1 signaling cooperates with papillomavirus oncogenes in transformation and generates resistance to apoptosis on matrix withdrawal through PKB/Akt. Virology 286: 23–30.
Reedijk M, Odorcic S, Chang L, Zhang H, Miller N, McCready DR et al. (2005). High-level coexpression of JAG1 and NOTCH1 is observed in human breast cancer and is associated with poor overall survival. Cancer Res 65: 8530–8537.
Reedijk M, Pinnaduwage D, Dickson BC, Mulligan AM, Zhang H, Bull SB et al. (2007). JAG1 expression is associated with a basal phenotype and recurrence in lymph node-negative breast cancer. Breast Cancer Res Treat; e-pub ahead of print.
Romond EH, Perez EA, Bryant J, Suman VJ, Geyer Jr CE, Davidson NE et al. (2005). Trastuzumab plus adjuvant chemotherapy for operable HER2-positive breast cancer. N Engl J Med 353: 1673–1684.
Ronchini C, Capobianco AJ . (2001). Induction of cyclin D1 transcription and CDK2 activity by Notch(ic): implication for cell cycle disruption in transformation by Notch(ic). Mol Cell Biol 21: 5925–5934.
Schroeter EH, Kisslinger JA, Kopan R . (1998). Notch-1 signalling requires ligand-induced proteolytic release of intracellular domain. Nature 393: 382–386.
Seidman AD, Fornier MN, Esteva FJ, Tan L, Kaptain S, Bach A et al. (2001). Weekly trastuzumab and paclitaxel therapy for metastatic breast cancer with analysis of efficacy by HER2 immunophenotype and gene amplification. J Clin Oncol 19: 2587–2595.
Shou J, Massarweh S, Osborne CK, Wakeling AE, Ali S, Weiss H et al. (2004). Mechanisms of tamoxifen resistance: increased estrogen receptor-HER2/neu cross-talk in ER/HER2-positive breast cancer. J Natl Cancer Inst 96: 926–935.
Sicinska E, Aifantis I, Le Cam L, Swat W, Borowski C, Yu Q et al. (2003). Requirement for cyclin D3 in lymphocyte development and T cell leukemias. Cancer Cell 4: 451–461.
Slamon DJ, Clark GM, Wong SG, Levin WJ, Ullrich A, McGuire WL . (1987). Human breast cancer: correlation of relapse and survival with amplification of the HER-2/neu oncogene. Science 235: 177–182.
Slamon DJ, Godolphin W, Jones LA, Holt JA, Wong SG, Keith DE et al. (1989). Studies of the HER-2/neu proto-oncogene in human breast and ovarian cancer. Science 244: 707–712.
Slamon DJ, Leyland-Jones B, Shak S, Fuchs H, Paton V, Bajamonde A et al. (2001). Use of chemotherapy plus a monoclonal antibody against HER2 for metastatic breast cancer that overexpresses HER2. N Engl J Med 344: 783–792.
Sliwkowski MX, Lofgren JA, Lewis GD, Hotaling TE, Fendly BM, Fox JA . (1999). Nonclinical studies addressing the mechanism of action of trastuzumab (Herceptin). Semin Oncol 26: 60–70.
Stylianou S, Clarke RB, Brennan K . (2006). Aberrant activation of notch signaling in human breast cancer. Cancer Res 66: 1517–1525.
Vogel CL, Cobleigh MA, Tripathy D, Gutheil JC, Harris LN, Fehrenbacher L et al. (2002). Efficacy and safety of trastuzumab as a single agent in first-line treatment of HER2-overexpressing metastatic breast cancer. J Clin Oncol 20: 719–726.
Vooijs M, Schroeter EH, Pan Y, Blandford M, Kopan R . (2004). Ectodomain shedding and intramembrane cleavage of mammalian Notch proteins is not regulated through oligomerization. J Biol Chem 279: 50864–50873.
Weber U, Eroglu C, Mlodzik M . (2003). Phospholipid membrane composition affects EGF receptor and Notch signaling through effects on endocytosis during Drosophila development. Dev Cell 5: 559–570.
Weijzen S, Rizzo P, Braid M, Vaishnav R, Jonkheer SM, Zlobin A et al. (2002). Activation of Notch-1 signaling maintains the neoplastic phenotype in human Ras-transformed cells. Nat Med 8: 979–986.
Weinmaster G . (1997). The ins and outs of notch signaling. Mol Cell Neurosci 9: 91–102.
Weng AP, Millholland JM, Yashiro-Ohtani Y, Arcangeli ML, Lau A, Wai C et al. (2006). c-Myc is an important direct target of Notch1 in T-cell acute lymphoblastic leukemia/lymphoma. Genes Dev 20: 2096–2109.
Xia W, Bacus S, Hegde P, Husain I, Strum J, Liu L et al. (2006). A model of acquired autoresistance to a potent ErbB2 tyrosine kinase inhibitor and a therapeutic strategy to prevent its onset in breast cancer. Proc Natl Acad Sci USA 103: 7795–7800.
Yarden Y . (2001). Biology of HER2 and its importance in breast cancer. Oncology 61 (Suppl 2): 1–13.
Yokoyama G, Fujii T, Ogo E, Yanaga H, Toh U, Yamaguchi M et al. (2005). Advanced chemoresistant breast cancer responding to multidisciplinary treatment with hyperthermia, radiotherapy, and intraarterial infusion. Int J Clin Oncol 10: 139–143.
Acknowledgements
We thank Dr Mien-Chie Hung (MD Anderson Cancer Center, Houston, TX) for MCF-7/Neo and MCF-7/HER2-18 cells. We are grateful to Dr Peter Strack (Merck Inc.) for the MRK-003 GSI. We are very thankful to Dr Rapheal Kopan (Washington University School of Medicine, Saint Louis, MO) for the full-length Notch-1 Renilla construct. This research was kindly supported by a grant no. 08-05 from the American Cancer Society, Illinois Division.
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Osipo, C., Patel, P., Rizzo, P. et al. ErbB-2 inhibition activates Notch-1 and sensitizes breast cancer cells to a γ-secretase inhibitor. Oncogene 27, 5019–5032 (2008). https://doi.org/10.1038/onc.2008.149
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DOI: https://doi.org/10.1038/onc.2008.149
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