Surgery combined with new adjuvant chemotherapy is the primary treatment for early stage invasive and advanced stage breast cancer. Growing evidence indicates that patients with ERα-positive breast cancer show poor response to chemotherapeutics. However, ERα-mediated drug-resistant mechanisms remain unclear.
Levels of WW domain-binding protein 2 (WBP2) and drug-resistant gene were determined by western blotting and RT-PCR, respectively. Cell viability was measured by preforming MTT assay. CD243 expression and apoptosis rate were evaluated by flow cytometry. Interactions of WBP2/ERα and ERα/MDR1 were detected by co-immunoprecipitation and chromatin immunoprecipitation (ChIP) assay, respectively.
There was an intrinsic link between WBP2 and ERα in drug-resistant cancer cells. Upregulation of WBP2 in MCF7 cells increased the chemoresistance to doxorubicin, while RNAi-mediated knockdown of WBP2 in MCF7/ADR cells sensitised the cancer cells to doxorubicin. Further investigation in in vitro and in vivo models demonstrated that WBP2 expression was directly correlated with MDR1, and WBP2 could directly modulate MDR1 transcription through binding to ERα, resulting in increased chemotherapy drug resistance.
Our finding provides a new mechanism for the chemotherapy response of ERα-positive breast tumours, and WBP2 might be a key molecule for developing new therapeutic strategies to treat chemoresistance in breast cancer patients.
Access optionsAccess options
Subscribe to Journal
Get full journal access for 1 year
only $32.88 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Note: This work is published under the standard license to publish agreement. After 12 months the work will become freely available and the license terms will switch to a Creative Commons Attribution 4.0 International (CC BY 4.0).
DeSantis, C. E. et al. Cancer treatment and survivorship statistics, 2014. CA Cancer J. Clin. 64, 252–271 (2014).
Regan, M. M. & Gelber, R. D. Predicting response to systemic treatments: learning from the past to plan for the future. Breast 14, 582–593 (2005).
Pruthi, S. et al. A multidisciplinary approach to the management of breast cancer, part 2: therapeutic considerations. Mayo Clin. Proc. 82, 1131–1140 (2007).
Sledge, G. W. et al. Phase III trial of doxorubicin, paclitaxel, and the combination of doxorubicin and paclitaxel as front-line chemotherapy for metastatic breast cancer: an intergroup trial (E1193). J. Clin. Oncol. 21, 588–592 (2003).
Ning, Y. Z. et al. ATP-binding cassette transporter enhances tolerance to DDT in tetrahymena. Sci. China Life Sci. 58, 297–304 (2015).
Zhang, J. T. Use of arrays to investigate the contribution of ATP-binding cassette transporters to drug resistance in cancer chemotherapy and prediction of chemosensitivity. Cell Res. 17, 311–323 (2007).
Choi, Y. H. & Yu, A. M. ABC transporters in multidrug resistance and pharmacokinetics, and strategies for drug development. Curr. Pharm. Des. 20, 793–807 (2014).
Yan, Y. et al. Rack1 mediates the interaction of P-glycoprotein with anxa2 and regulates migration and invasion of multidrug-resistant breast cancer cells. Int. J. Mol. Sci. 17, E1718 (2016).
Levi, M., Brunetti, B., Sarli, G. & Benazzi, C. Immunohistochemical expression of P-glycoprotein and breast cancer resistance protein in canine mammary hyperplasia, neoplasia and supporting stroma. J. Comp. Pathol. 155, 277–285 (2016).
Puhalla, S., Bhattacharya, S. & Davidson, N. E. Hormonal therapy in breast cancer: a model disease for the personalization of cancer care. Mol. Oncol. 6, 222–236 (2012).
Lips, E. H. et al. Neoadjuvant chemotherapy in ER+ HER2-breast cancer: response prediction based on immunohistochemical and molecular characteristics. Breast Cancer Res. Tr. 131, 827–836 (2012).
Bailey, S. T., Shin, H. J., Westerling, T., Liu, X. S. & Brown, M. Estrogen receptor prevents p53-dependent apoptosis in breast cancer. Proc. Natl Acad. Sci. USA 109, 18060–18065 (2012).
Sui, M., Huang, Y., Park, B. H., Davidson, N. E. & Fan, W. Estrogen receptor alpha mediates breast cancer cell resistance to paclitaxel through inhibition of apoptotic cell death. Cancer Res. 67, 5337–5344 (2007).
Chang, J. J., Sui, M. H. & Fan, W. M. Estrogen receptor alpha attenuates therapeutic efficacy of paclitaxel on breast xenograft tumors. Breast Cancer Res. Tr. 134, 969–980 (2012).
Shi, J. F. et al. ER alpha directly activated the MDR1 transcription to increase paclitaxel-resistance of ER alpha-positive breast cancer cells in vitro and in vivo. Int. J. Biochem. Cell B 53, 35–45 (2014).
Si, X. X. et al. ER alpha propelled aberrant global DNA hypermethylation by activating the DNMT1 gene to enhance anticancer drug resistance in human breast cancer cells. Oncotarget 7, 20966–20980 (2016).
Shi, J. F. et al. ER alpha positively regulated DNMT1 expression by binding to the gene promoter region in human breast cancer MCF-7 cells. Biochem. Biophys. Res. Commun. 427, 47–53 (2012).
Sui, M., Zhang, H. & Fan, W. The role of estrogen and estrogen receptors in chemoresistance. Curr. Med. Chem. 18, 4674–4683 (2011).
Ross-Innes, C. S. et al. Differential oestrogen receptor binding is associated with clinical outcome in breast cancer. Nature 481, 389–U177 (2012).
Chan, S. W. et al. WW domain-mediated interaction with Wbp2 is important for the oncogenic property of TAZ. Oncogene 30, 600–610 (2011).
Chen, S. et al. WW domain-binding protein 2: an adaptor protein closely linked to the development of breast cancer. Mol. Cancer. 16, 128 (2017).
Buffa, L., Saeed, A. M. & Nawaz, Z. Molecular mechanism of WW-domain binding protein-2 coactivation function in estrogen receptor signaling. IUBMB Life 65, 76–84 (2013).
McDonald, C. B. et al. Biophysical basis of the binding of WWOX tumor suppressor to WBP1 and WBP2 adaptors. J. Mol. Biol. 422, 58–74 (2012).
Lim, S. K., Orhant-Prioux, M., Toy, W., Tan, K. Y. & Lim, Y. P. Tyrosine phosphorylation of transcriptional coactivator WW-domain binding protein 2 regulates estrogen receptor alpha function in breast cancer via the Wnt pathway. FASEB J. 25, 3004–3018 (2011).
Lim, S. K. et al. Wnt signaling promotes breast cancer by blocking ITCH-mediated degradation of YAP/TAZ transcriptional coactivator WBP2. Cancer Res. 76, 6278–6289 (2016).
Chen, S. et al. Deletion of Gab2 in mice protects against hepatic steatosis and steatohepatitis: a novel therapeutic target for fatty liver disease. J. Mol. Cell Biol. 8, 492–504 (2016).
Mukhopadhyay, A., Deplancke, B., Walhout, A. J. M. & Tissenbaum, H. A. Chromatin immunoprecipitation (ChIP) coupled to detection by quantitative real-time PCR to study transcription factor binding to DNA in Caenorhabditis elegans. Nat. Protoc. 3, 698–709 (2008).
Cheng, J. H. et al. Gab2 mediates hepatocellular carcinogenesis by integrating multiple signaling pathways. FASEB J. 31, 5530–553 (2017).
Ren, Y. Q., Wang, H. J., Zhang, Y. Q. & Liu, Y. B. WBP2 modulates G1/S transition in ER plus breast cancer cells and is a direct target of miR-206. Cancer Chemoth. Pharm. 79, 1003–1011 (2017).
Nourashrafeddin, S., Aarabi, M., Modarressi, M. H., Rahmati, M. & Nouri, M. The revaluation of WBP2NL-related genes expression in breast cancer. Pathol. Oncol. Res. 21, 293–300 (2015).
Salvat, C., Wang, G. L., Dastur, A., Lyon, N. & Huibregtse, J. M. The-4 phenylalanine is required for substrate ubiquitination catalyzed by HECT ubiquitin ligases. J. Biol. Chem. 279, 18935–18943 (2004).
Nitsch, R., Di Palma, T., Mascia, A. & Zannini, M. WBP-2, a WW domain binding protein, interacts with the thyroid-specific transcription factor Pax8. Biochem. J. 377, 553–560 (2004).
Chen, S. et al. WW domain-binding protein 2 acts as an oncogene by modulating the activity of the glycolytic enzyme ENO1 in glioma. Cell Death Dis. 9, 347 (2018).
Zhang, X., Milton, C. C., Poon, C. L. C., Hong, W. & Harvey, K. F. Wbp2 cooperates with Yorkie to drive tissue growth downstream of the Salvador-Warts-Hippo pathway. Cell Death Differ. 18, 1346–1355 (2011).
Aller, S. G. et al. Structure of P-glycoprotein reveals a molecular basis for poly-specific drug binding. Science 323, 1718–1722 (2009).
Colabufo, N. A. et al. Substrates, inhibitors and activators of P-glycoprotein: candidates for radiolabeling and imaging perspectives. Curr. Top. Med. Chem. 10, 1703–1714 (2010).
Wesolowska, O. Interaction of phenothiazines, stilbenes and flavonoids with multidrug resistance-associated transporters, P-glycoprotein and MRP1. Acta Biochim. Pol. 58, 433–448 (2011).
Xia, W. H. et al. Celecoxib enhanced the sensitivity of cancer cells to anticancer drugs by inhibition of the expression of P-glycoprotein through a COX-2-independent manner. J. Cell Biochem. 108, 181–194 (2009).
Jin, W. et al. UHRF1 inhibits MDR1 gene transcription and sensitizes breast cancer cells to anticancer drugs. Breast Cancer Res. Tr. 124, 39–48 (2010).
Barone, I., Brusco, L. & Fuqua, S. A. W. Estrogen receptor mutations and changes in downstream gene expression and signaling. Clin. Cancer Res. 16, 2702–2708 (2010).
Wang, C. Y. et al. Estrogen induces c-myc gene expression via an upstream enhancer activated by the estrogen receptor and the AP-1 transcription factor. Mol. Endocrinol. 25, 1527–1538 (2011).
Pradhan, M., Bembinster, L. A., Baumgarten, S. C. & Frasor, J. 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 (2010).
Labialle, S., Gayet, L., Marthinet, E., Rigal, D. & Baggetto, L. G. Transcriptional regulators of the human multidrug resistance 1 gene: recent views. Biochem. Pharmacol. 64, 943–948 (2002).
Huo, H. R., Magro, P. G., Pietsch, E. C., Patel, B. B. & Scotto, K. W. Histone methyltransferase MLL1 regulates MDR1 transcription and chemoresistance. Cancer Res. 70, 8726–8735 (2010).
Cornwell, M. M. & Smith, D. E. SP1 activates the MDR1 promoter through one of two distinct G-rich regions that modulate promoter activity. J. Biol. Chem. 268, 19505–19511 (1993).
We thank LetPub (www.letpub.com) for its linguistic assistance in this manuscript.
C.-M.T. and Z.M. conceived the idea; C.-M.T. and S.C. drafted the manuscript; S.C. and H.W. designed and performed the experiments; S.C. and J.Y. analysed the data and designed the figures; Z.L. and Y.Y. contributed to western blotting assay; Q.W. provided the clinical samples of breast cancer. All authors discussed the results and edited this manuscript.