Synergy of leptin/STAT3 with HER2 receptor induces tamoxifen resistance in breast cancer cells through regulation of apoptosis-related genes
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Tamoxifen is a major treatment modality for estrogen receptor positive breast cancer, but the occurrence of resistance remains a problem. Recently, obesity-related leptin has been found to interfere with tamoxifen in breast cancer MCF-7 cells. In the present study we investigated the effect of leptin on three tamoxifen-treated breast cancer cell types (i.e., MDA-MB-231, MCF-7 and MCF-7/HER2).
The effect of tamoxifen/leptin treatment was evaluated using a MTT cell viability assay. mRNA expression was assessed by real time PCR and protein expression by Western blotting. WWOX, Survivin and BCL2 gene promoter activities were evaluated by chromatin immunoprecipitation.
Cell viability assays revealed that estrogen receptor negative MDA-MB-231 cells were resistant, that estrogen receptor positive MCF-7 cells were sensitive and that MCF-7/HER2 cells were relatively resistant to tamoxifen, while leptin co-administration ‘rescued’ MCF-7 and, especially, MCF-7/HER2 cells from the anti-proliferative effect of tamoxifen. The cell lines also exhibited a different phosphorylation status of STAT3, a transcription factor that is activated by the obesity related leptin receptor b (Ob-Rb). Most importantly, chromatin immunoprecipitation assays revealed differential STAT3 binding to the anti-apoptotic BCL2 and pro-apoptotic WWOX gene promoters in MCF-7 and MCF-7/HER2 cells, leading to concomitant modifications of its mRNA/protein expression levels, thus providing a selective advantage to HER2 over-expressing MCF-7/HER2 cells after treatment with tamoxifen and tamoxifen plus leptin.
Our study provides novel evidence indicating that synergy between the leptin/Ob-Rb/STAT3 signalling pathway and the HER2 receptor protects tamoxifen-treated HER2 over-expressing cells from the inhibitory effect of tamoxifen through differential regulation of apoptosis-related genes.
KeywordsTamoxifen Leptin HER2 receptor STAT3 Breast cancer BCL2 WWOX
Conflict of interest
The authors have no conflict of interest to report with respect to this work.
- 2.B. Pula, M. Olbromski, A. Wojnar, A. Gomulkiewicz, W. Witkiewicz, M. Ugorski, P. Dziegiel, M. Podhorska-Okolow, Impact of SOX18 expression in cancer cells and vessels on the outcome of invasive ductal breast carcinoma. Cell. Oncol. 36, 469–483 (2013)Google Scholar
- 4.J.S. de Groot, X. Pan, J. Meeldijk, E. van der Wall, P.J. van Diest, C.B. Moelans, Validation of DNA promoter hypermethylation biomarkers in breast cancer–a short report. Cell. Oncol. 37, 297–303 (2014)Google Scholar
- 5.S. Loi, B. Haibe-Kains, C. Desmedt, P. Wirapati, F. Lallemand, A.M. Tutt, C. Gillet, P. Ellis, K. Ryder, J.F. Reid, M.G. Daidone, M.A. Pierotti, E.M. Berns, M.P. Jansen, J.A. Foekens, M. Delorenzi, G. Bontempi, M.J. Piccart, C. Sotiriou, Predicting prognosis using molecular profiling in estrogen receptor-positive breast cancer treated with tamoxifen. BMC Genomics 9, 239 (2008)CrossRefPubMedCentralPubMedGoogle Scholar
- 6.A. Halon, P. Donizy, P. Surowiak, R. Matkowski, ERM/Rho protein expression in ductal breast cancer: a 15 year follow-up. Cell. Oncol. 36, 181–190 (2013)Google Scholar
- 7.S. Tabarestani, S.M. Ghaderian, H. Rezvani, R. Mirfakhraie, A. Ebrahimi, H. Attarian, J. Rafat, M. Ghadyani, H.A. Alavi, N. Kamalian, A. Rakhsha, E. Azargashb, Prognostic and predictive value of copy number alterations in invasive breast cancer as determined by multiplex ligation-dependent probe amplification. Cell. Oncol. 37, 107–118 (2014)Google Scholar
- 12.S.H. Kim, A. Nagalingam, N.K. Saxena, S.V. Singh, D. Sharma, Benzyl isothiocyanate inhibits oncogenic actions of leptin in human breast cancer cells by suppressing activation of signal transducer and activator of transcription 3. Carcinogenesis 32, 359–367 (2011)CrossRefPubMedCentralPubMedGoogle Scholar
- 21.L.A. Tartaglia, M. Dembski, X. Weng, N. Deng, J. Culpepper, R. Devos, G.J. Richards, L.A. Campfield, F.T. Clark, J. Deeds, C. Muir, S. Sanker, A. Moriarty, K.J. Moore, J.S. Smutko, G.G. Mays, E.A. Wool, C.A. Monroe, R.I. Tepper, Identification and expression cloning of a leptin receptor, OB-R. Cell 83, 1263–1271 (1995)CrossRefPubMedGoogle Scholar
- 28.R. Devos, Y. Guisez, J. Van der Heyden, D.W. White, M. Kalai, M. Fountoulakis, G. Plaetinck, Ligand-independent dimerization of the extracellular domain of the leptin receptor and determination of the stoichiometry of leptin binding. J. Biol. Chem. 272, 18304–18310 (1997)CrossRefPubMedGoogle Scholar
- 30.Q. Gao, M.J. Wolfgang, S. Neschen, K. Morino, T.L. Horvath, G.I. Shulman, X.Y. Fu, Disruption of neural signal transducer and activator of transcription 3 causes obesity, diabetes, infertility, and thermal dysregulation. Proc. Natl. Acad. Sci. U. S. A. 101, 4661–4666 (2004)CrossRefPubMedCentralPubMedGoogle Scholar
- 31.H. Inoue, W. Ogawa, M. Ozaki, S. Haga, M. Matsumoto, K. Furukawa, N. Hashimoto, Y. Kido, T. Mori, H. Sakaue, K. Teshigawara, S. Jin, H. Iguchi, R. Hiramatsu, D. LeRoith, K. Takeda, S. Akira, M. Kasuga, Role of STAT-3 in regulation of hepatic gluconeogenic genes and carbohydrate metabolism in vivo. Nat. Med. 10, 168–174 (2004)CrossRefPubMedGoogle Scholar
- 33.J. Abdulghani, L. Gu, A. Dagvadorj, J. Lutz, B. Leiby, G. Bonuccelli, M.P. Lisanti, T. Zellweger, K. Alanen, T. Mirtti, T. Visakorpi, L. Bubendorf, M.T. Nevalainen, Stat3 promotes metastatic progression of prostate cancer. Am. J. Pathol. 172, 1717–1728 (2008)CrossRefPubMedCentralPubMedGoogle Scholar
- 34.R. Catlett-Falcone, T.H. Landowski, M.M. Oshiro, J. Turkson, A. Levitzki, R. Savino, G. Ciliberto, L. Moscinski, J.L. Fernandez-Luna, G. Nunez, W.S. Dalton, R. Jove, Constitutive activation of Stat3 signaling confers resistance to apoptosis in human U266 myeloma cells. Immunity 10, 105–115 (1999)CrossRefPubMedGoogle Scholar
- 36.T. Gritsko, A. Williams, J. Turkson, S. Kaneko, T. Bowman, M. Huang, S. Nam, I. Eweis, N. Diaz, D. Sullivan, S. Yoder, S. Enkemann, S. Eschrich, J.H. Lee, C.A. Beam, J. Cheng, S. Minton, C.A. Muro-Cacho, R. Jove, Persistent activation of stat3 signaling induces survivin gene expression and confers resistance to apoptosis in human breast cancer cells. Clin. Cancer Res. 12, 11–19 (2006)CrossRefPubMedGoogle Scholar
- 37.B.B. Aggarwal, G. Sethi, K.S. Ahn, S.K. Sandur, M.K. Pandey, A.B. Kunnumakkara, B. Sung, H. Ichikawa, Targeting signal-transducer-and-activator-of-transcription-3 for prevention and therapy of cancer: modern target but ancient solution. Ann. N. Y. Acad. Sci. 1091, 151–169 (2006)Google Scholar
- 40.E. Fiorio, A. Mercanti, M. Terrasi, R. Micciolo, A. Remo, A. Auriemma, A. Molino, V. Parolin, B. Di Stefano, F. Bonetti, A. Giordano, G.L. Cetto, E. Surmacz, Leptin/HER2 crosstalk in breast cancer: in vitro study and preliminary in vivo analysis. BMC Cancer 8, 305 (2008)CrossRefPubMedCentralPubMedGoogle Scholar
- 41.X. Chen, X. Zha, W. Chen, T. Zhu, J. Qiu, O.D. Roe, J. Li, Z. Wang, Y. Yin, Leptin attenuates the anti-estrogen effect of tamoxifen in breast cancer. Biomed. Pharmacother. 67, 22–30 (2012)Google Scholar
- 43.P.H. Lahmann, K. Hoffmann, N. Allen, C.H. van Gils, K.T. Khaw, B. Tehard, F. Berrino, A. Tjonneland, J. Bigaard, A. Olsen, K. Overvad, F. Clavel-Chapelon, G. Nagel, H. Boeing, D. Trichopoulos, G. Economou, G. Bellos, D. Palli, R. Tumino, S. Panico, C. Sacerdote, V. Krogh, P.H. Peeters, H.B. Bueno-de-Mesquita, E. Lund, E. Ardanaz, P. Amiano, G. Pera, J.R. Quiros, C. Martinez, M.J. Tormo, E. Wirfalt, G. Berglund, G. Hallmans, T.J. Key, G. Reeves, S. Bingham, T. Norat, C. Biessy, R. Kaaks, E. Riboli, Body size and breast cancer risk: findings from the European Prospective Investigation into Cancer And Nutrition (EPIC). Int. J. Cancer 111, 762–771 (2004)CrossRefPubMedGoogle Scholar
- 50.A. Valle, J. Sastre-Serra, J. Oliver, P. Roca, Chronic leptin treatment sensitizes MCF-7 breast cancer cells to estrogen. Cell Physiol. Biochem. 28, 823–32 (2011)Google Scholar
- 51.C. Carlomagno, F. Perrone, C. Gallo, M. De Laurentiis, R. Lauria, A. Morabito, G. Pettinato, L. Panico, A. D’Antonio, A.R. Bianco, S. De, Placido, c-erb B2 overexpression decreases the benefit of adjuvant tamoxifen in early-stage breast cancer without axillary lymph node metastases. J. Clin. Oncol. 14, 2702–8 (1996)PubMedGoogle Scholar
- 52.C.K. Osborne, V. Bardou, T.A. Hopp, G.C. Chamness, S.G. Hilsenbeck, S.A. Fuqua, J. Wong, D.C. Allred, G.M. Clark, R. Schiff, 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 (2003)CrossRefPubMedGoogle Scholar
- 53.I.E. Smith, M. Dowsett, S.R. Ebbs, J.M. Dixon, A. Skene, J.U. Blohmer, S.E. Ashley, S. Francis, I. Boeddinghaus, G. Walsh, Neoadjuvant treatment of postmenopausal breast cancer with anastrozole, tamoxifen, or both in combination: the Immediate Preoperative Anastrozole, Tamoxifen, or Combined with Tamoxifen (IMPACT) multicenter double-blind randomized trial. J. Clin. Oncol. 23, 5108–5116 (2005)CrossRefPubMedGoogle Scholar
- 56.P.K. Epling-Burnette, J.H. Liu, R. Catlett-Falcone, J. Turkson, M. Oshiro, R. Kothapalli, Y. Li, J.M. Wang, H.F. Yang-Yen, J. Karras, R. Jove, T.P. Loughran Jr., Inhibition of STAT3 signaling leads to apoptosis of leukemic large granular lymphocytes and decreased Mcl-1 expression. J. Clin. Invest. 107, 351–362 (2001)CrossRefPubMedCentralPubMedGoogle Scholar
- 57.S. Alas, B. Bonavida, Rituximab inactivates signal transducer and activation of transcription 3 (STAT3) activity in B-non-Hodgkin’s lymphoma through inhibition of the interleukin 10 autocrine/paracrine loop and results in down-regulation of Bcl-2 and sensitization to cytotoxic drugs. Cancer Res. 61, 5137–5144 (2001)PubMedGoogle Scholar
- 59.N. Diaz, S. Minton, C. Cox, T. Bowman, T. Gritsko, R. Garcia, I. Eweis, M. Wloch, S. Livingston, E. Seijo, A. Cantor, J.H. Lee, C.A. Beam, D. Sullivan, R. Jove, C.A. Muro-Cacho, Activation of stat3 in primary tumors from high-risk breast cancer patients is associated with elevated levels of activated SRC and survivin expression. Clin. Cancer Res. 12, 20–28 (2006)CrossRefPubMedGoogle Scholar
- 66.R.I. Aqeilan, Y. Pekarsky, J.J. Herrero, A. Palamarchuk, J. Letofsky, T. Druck, F. Trapasso, S.Y. Han, G. Melino, K. Huebner, C.M. Croce, Functional association between Wwox tumor suppressor protein and p73, a p53 homolog. Proc. Natl. Acad. Sci. U. S. A. 101, 4401–4406 (2004)CrossRefPubMedCentralPubMedGoogle Scholar
- 70.D. Trivigno, F. Essmann, S.M. Huber, J. Rudner, Deubiquitinase USP9x confers radioresistance through stabilization of Mcl-1. Neoplasia, 14, 893–904 (2012)Google Scholar