{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,24]],"date-time":"2026-08-24T15:39:10Z","timestamp":1787585950621,"version":"build-2736575974"},"reference-count":102,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2024,6,21]],"date-time":"2024-06-21T00:00:00Z","timestamp":1718928000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Conselho Nacional de Desenvolvimento Cient\u00edfico e Tecnol\u00f3gico (CNPq)"},{"name":"Funda\u00e7\u00e3o Carlos Chagas de Amparo \u00e0 Pesquisa do Estado do Rio de Janeiro (FAPERJ)"},{"name":"Programa de Objetivos e Metas\u2014Fiocruz (POM\/Fiocruz)"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biology"],"abstract":"<jats:p>Breast cancer is the most prevalent cancer among women worldwide. Therapeutic strategies to control tumors and metastasis are still challenging. Three-dimensional (3D) spheroid-type systems more accurately replicate the features of tumors in vivo, working as a better platform for performing therapeutic response analysis. This work aimed to characterize the epithelial\u2013mesenchymal transition and doxorubicin (dox) response in a mammary tumor spheroid (MTS) model. We evaluated the doxorubicin treatment effect on MCF-7 spheroid diameter, cell viability, death, migration and proteins involved in the epithelial\u2013mesenchymal transition (EMT) process. Spheroids were also produced from tumors formed from 4T1 and 67NR cell lines. MTSs mimicked avascular tumor characteristics, exhibited adherens junction proteins and independently produced their own extracellular matrix. Our spheroid model supports the 3D culturing of cells isolated from mice mammary tumors. Through the migration assay, we verified a reduction in E-cadherin expression and an increase in vimentin expression as the cells became more distant from spheroids. Dox promoted cytotoxicity in MTSs and inhibited cell migration and the EMT process. These results suggest, for the first time, that this model reproduces aspects of the EMT process and describes the potential of dox in inhibiting the metastatic process, which can be further explored.<\/jats:p>","DOI":"10.3390\/biology13070463","type":"journal-article","created":{"date-parts":[[2024,6,21]],"date-time":"2024-06-21T12:27:33Z","timestamp":1718972853000},"page":"463","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["Spheroid Model of Mammary Tumor Cells: Epithelial\u2013Mesenchymal Transition and Doxorubicin Response"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5242-9541","authenticated-orcid":false,"given":"Laura Lacerda","family":"Coelho","sequence":"first","affiliation":[{"name":"Laboratory of Innovations in Therapies, Education and Bioproducts, Oswaldo Cruz Institute (IOC), Oswaldo Cruz Foundation (Fiocruz), Rio de Janeiro 21040-900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matheus Menezes","family":"Vianna","sequence":"additional","affiliation":[{"name":"Laboratory of Innovations in Therapies, Education and Bioproducts, Oswaldo Cruz Institute (IOC), Oswaldo Cruz Foundation (Fiocruz), Rio de Janeiro 21040-900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Debora Moraes","family":"da Silva","sequence":"additional","affiliation":[{"name":"Laboratory of Innovations in Therapies, Education and Bioproducts, Oswaldo Cruz Institute (IOC), Oswaldo Cruz Foundation (Fiocruz), Rio de Janeiro 21040-900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6862-7008","authenticated-orcid":false,"given":"Beatriz Matheus de Souza","family":"Gonzaga","sequence":"additional","affiliation":[{"name":"Laboratory of Innovations in Therapies, Education and Bioproducts, Oswaldo Cruz Institute (IOC), Oswaldo Cruz Foundation (Fiocruz), Rio de Janeiro 21040-900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Roberto Rodrigues","family":"Ferreira","sequence":"additional","affiliation":[{"name":"Laboratory of Innovations in Therapies, Education and Bioproducts, Oswaldo Cruz Institute (IOC), Oswaldo Cruz Foundation (Fiocruz), Rio de Janeiro 21040-900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ana Carolina","family":"Monteiro","sequence":"additional","affiliation":[{"name":"Laboratory of Osteo and Tumor Immunology, Department of Immunobiology, Fluminense Federal University (UFF), Rio de Janeiro 24020-150, Brazil"},{"name":"Thymus Research Laboratory, Oswaldo Cruz Institute (IOC), Oswaldo Cruz Foundation (Fiocruz), Rio de Janeiro 21040-900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adriana Cesar","family":"Bonomo","sequence":"additional","affiliation":[{"name":"Thymus Research Laboratory, Oswaldo Cruz Institute (IOC), Oswaldo Cruz Foundation (Fiocruz), Rio de Janeiro 21040-900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pedro Paulo de Abreu","family":"Manso","sequence":"additional","affiliation":[{"name":"Laboratory of Pathology, Oswaldo Cruz Institute (IOC), Oswaldo Cruz Foundation (Fiocruz), Rio de Janeiro 21040-900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7053-0053","authenticated-orcid":false,"given":"Marcelo Alex","family":"de Carvalho","sequence":"additional","affiliation":[{"name":"Research Center (CPQ), National Cancer Institute (INCA), Rio de Janeiro 20231-050, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fernando Regla","family":"Vargas","sequence":"additional","affiliation":[{"name":"Laboratory of Epidemiology of Congenital Malformations, Oswaldo Cruz Institute (IOC), Oswaldo Cruz Foundation (Fiocruz), Rio de Janeiro 21040-900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luciana Ribeiro","family":"Garzoni","sequence":"additional","affiliation":[{"name":"Laboratory of Innovations in Therapies, Education and Bioproducts, Oswaldo Cruz Institute (IOC), Oswaldo Cruz Foundation (Fiocruz), Rio de Janeiro 21040-900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,6,21]]},"reference":[{"key":"ref_1","unstructured":"WHO (2024, February 01). Breast Cancer. Available online: https:\/\/www.who.int\/news-room\/fact-sheets\/detail\/breast-cancer."},{"key":"ref_2","first-page":"43","article-title":"Breast cancer in Brazil: Epidemiology and treatment challenges","volume":"7","author":"Cecilio","year":"2015","journal-title":"Breast Cancer"},{"key":"ref_3","unstructured":"(2023, October 13). BCNA. Available online: https:\/\/www.bcna.org.au\/resource-hub\/articles\/types-of-breast-cancer\/."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"e95","DOI":"10.1016\/S1470-2045(11)70323-0","article-title":"Breast cancer in Brazil: Present status and future goals","volume":"13","author":"Lee","year":"2012","journal-title":"Lancet Oncol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1016\/S0092-8674(00)81683-9","article-title":"The hallmarks of cancer","volume":"100","author":"Hanahan","year":"2000","journal-title":"Cell"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"646","DOI":"10.1016\/j.cell.2011.02.013","article-title":"Hallmarks of cancer: The next generation","volume":"144","author":"Hanahan","year":"2011","journal-title":"Cell"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1158\/2159-8290.CD-21-1059","article-title":"Hallmarks of Cancer: New Dimensions","volume":"12","author":"Hanahan","year":"2022","journal-title":"Cancer Discov."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"226","DOI":"10.1186\/bcr3037","article-title":"Breast cancer epithelial-to-mesenchymal transition: Examining the functional consequences of plasticity","volume":"13","author":"Drasin","year":"2011","journal-title":"Breast Cancer Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1186\/s40169-015-0048-3","article-title":"EMT and tumor metastasis","volume":"4","author":"Heerboth","year":"2015","journal-title":"Clin. Transl. Med."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Kang, E., Seo, J., Yoon, H., and Cho, S. (2021). The Post-Translational Regulation of Epithelial-Mesenchymal Transition-Inducing Transcription Factors in Cancer Metastasis. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22073591"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1615\/CritRevOncog.v18.i1-2.40","article-title":"On the origin of cancer metastasis","volume":"18","author":"Seyfried","year":"2013","journal-title":"Crit. Rev. Oncog."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"6958","DOI":"10.1038\/onc.2008.346","article-title":"Transcriptional regulation of cell polarity in EMT and cancer","volume":"27","author":"Portillo","year":"2008","journal-title":"Oncogene"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Zhitnyak, I.Y., Rubtsova, S.N., Litovka, N.I., and Gloushankova, N.A. (2020). Early Events in Actin Cytoskeleton Dynamics and E-Cadherin-Mediated Cell-Cell Adhesion during Epithelial-Mesenchymal Transition. Cells, 9.","DOI":"10.3390\/cells9030578"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"764","DOI":"10.1016\/j.tcb.2020.07.003","article-title":"EMT, MET, Plasticity, and Tumor Metastasis","volume":"30","author":"Bakir","year":"2020","journal-title":"Trends Cell Biol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"3033","DOI":"10.1007\/s00018-011-0735-1","article-title":"Vimentin in cancer and its potential as a molecular target for cancer therapy","volume":"68","author":"Satelli","year":"2011","journal-title":"Cell Mol. Life Sci."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"14483","DOI":"10.1038\/ncomms14483","article-title":"Loss of ER\u03b1 induces amoeboid-like migration of breast cancer cells by downregulating vinculin","volume":"8","author":"Gao","year":"2017","journal-title":"Nat. Commun."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"dmm034330","DOI":"10.1242\/dmm.034330","article-title":"Intravital microscopy of collective invasion plasticity in breast cancer","volume":"11","author":"Ilina","year":"2018","journal-title":"Dis. Model. Mech."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"421","DOI":"10.1007\/s10585-017-9858-6","article-title":"Collective invasion in ductal and lobular breast cancer associates with distant metastasis","volume":"34","author":"Khalil","year":"2017","journal-title":"Clin. Exp. Metastasis"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/j.devcel.2010.06.013","article-title":"Cancer Dissemination-Lessons from Leukocytes","volume":"19","author":"Madsen","year":"2010","journal-title":"Dev. Cell"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1111\/cge.12381","article-title":"Personalized therapy for breast cancer","volume":"86","author":"Schwartz","year":"2014","journal-title":"Clin. Genet."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"a001578","DOI":"10.1101\/mcs.a001578","article-title":"Personalized cancer therapy- leveraging a knowledge base for clinical decision-making","volume":"4","author":"Dumbrava","year":"2018","journal-title":"Cold Spring Harb. Mol. Case Stud."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"3797","DOI":"10.1172\/JCI57152","article-title":"Targeted therapies for breast cancer","volume":"121","author":"Higgins","year":"2011","journal-title":"J. Clin. Investig."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"120","DOI":"10.5306\/wjco.v8.i2.120","article-title":"Targeted therapies in breast cancer: New challenges to fight against resistance","volume":"8","author":"Masoud","year":"2017","journal-title":"World J. Clin. Oncol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"140","DOI":"10.1186\/s13058-015-0648-2","article-title":"The benefit of HER2-targeted therapies on overall survival of patients with metastatic HER2-positive breast cancer\u2014A systematic review","volume":"17","author":"Mendes","year":"2015","journal-title":"Breast Cancer Res."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1038\/s41591-020-01195-4","article-title":"Targeting metastatic cancer","volume":"27","author":"Ganesh","year":"2021","journal-title":"Nat. Med."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1065","DOI":"10.4252\/wjsc.v11.i12.1065","article-title":"Three-dimensional cell culture systems as an in vitro platform for cancer and stem cell modeling","volume":"11","author":"Chaicharoenaudomrung","year":"2019","journal-title":"World J. Stem Cells"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"939","DOI":"10.1177\/1535370216643772","article-title":"Liquid-based three-dimensional tumor models for cancer research and drug discovery","volume":"241","author":"Ham","year":"2016","journal-title":"Exp. Biol. Med."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Urz\u00ec, O., Gasparro, R., Costanzo, E., De Luca, A., Giavaresi, G., Fontana, S., and Alessandro, R. (2023). Three-Dimensional Cell Cultures: The Bridge between In Vitro and In Vivo Models. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms241512046"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"311","DOI":"10.1016\/j.semcancer.2008.03.013","article-title":"Unraveling the microenvironmental influences on the normal mammary gland and breast cancer","volume":"18","author":"Weigelt","year":"2008","journal-title":"Semin. Cancer Biol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"309","DOI":"10.1038\/nprot.2008.226","article-title":"Spheroid-based drug screen: Considerations and practical approach","volume":"4","author":"Friedrich","year":"2009","journal-title":"Nat. Protoc."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"14167","DOI":"10.1038\/s41598-017-14326-8","article-title":"Methods to Evaluate Cell Growth, Viability, and Response to Treatment in a Tissue Engineered Breast Cancer Model","volume":"7","author":"Goliwas","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_32","first-page":"1384","article-title":"Inhibiting ULK1 kinase decreases autophagy and cell viability in high-grade serous ovarian cancer spheroids","volume":"10","author":"Singha","year":"2020","journal-title":"Am. J. Cancer Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1007\/978-1-62703-125-7_13","article-title":"Three-dimensional culture of human breast epithelial cells: The how and the why","volume":"945","author":"Vidi","year":"2013","journal-title":"Methods Mol. Biol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1007\/s00441-004-1064-6","article-title":"Method for the generation and cultivation of functional three-dimensional mammary constructs without exogenous extracellular matrix","volume":"320","author":"Timmins","year":"2005","journal-title":"Cell Tissue Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1007\/BF00196343","article-title":"Distributions of oxygen, nutrient, and metabolic waste concentrations in multicellular spheroids and their dependence on spheroid parameters","volume":"19","author":"Groebe","year":"1991","journal-title":"Eur. Biophys. J."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"192","DOI":"10.1016\/j.jconrel.2012.04.045","article-title":"Opportunities and challenges for use of tumor spheroids as models to test drug delivery and efficacy","volume":"164","author":"Mehta","year":"2012","journal-title":"J. Control Release"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"819","DOI":"10.1517\/17460441.2012.708334","article-title":"The multicellular tumor spheroid model for high-throughput cancer drug discovery","volume":"7","author":"LaBarbera","year":"2012","journal-title":"Expert. Opin. Drug Discov."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"101621","DOI":"10.1016\/j.isci.2020.101621","article-title":"A 3D Bioprinter Specifically Designed for the High-Throughput Production of Matrix-Embedded Multicellular Spheroids","volume":"23","author":"Utama","year":"2020","journal-title":"IScience"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"4561","DOI":"10.1021\/acs.chemrev.0c00752","article-title":"High-Throughput Methods in the Discovery and Study of Biomaterials and Materiobiology","volume":"121","author":"Yang","year":"2021","journal-title":"Chem. Rev."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/j.mrrev.2012.08.001","article-title":"In vitro cell migration and invasion assays","volume":"752","author":"Kramer","year":"2013","journal-title":"Mutat. Res."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"e202000942","DOI":"10.26508\/lsa.202000942","article-title":"Extracellular matrix mediates moruloid-blastuloid morphodynamics in malignant ovarian spheroids","volume":"4","author":"Langthasa","year":"2021","journal-title":"Life Sci. Alliance"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"6816","DOI":"10.1038\/srep06816","article-title":"Reversal of anchorage-independent multicellular spheroid into a monolayer mimics a metastatic model","volume":"4","author":"Kunjithapatham","year":"2014","journal-title":"Sci. Rep."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"de Barros, A.P., Takiya, C.M., Garzoni, L.R., Leal-Ferreira, M.L., Dutra, H.S., Chiarini, L.B., Meirelles, M.N., Borojevic, R., and Rossi, M.I. (2010). Osteoblasts and bone marrow mesenchymal stromal cells control hematopoietic stem cell migration and proliferation in 3D in vitro model. PLoS ONE, 5.","DOI":"10.1371\/journal.pone.0009093"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"906","DOI":"10.1086\/528373","article-title":"Fibrosis and hypertrophy induced by Trypanosoma cruzi in a three-dimensional cardiomyocyte-culture system","volume":"197","author":"Garzoni","year":"2008","journal-title":"J. Infect. Dis."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"3406","DOI":"10.1016\/j.yexcr.2009.09.016","article-title":"Dissecting coronary angiogenesis: 3D co-culture of cardiomyocytes with endothelial or mesenchymal cells","volume":"315","author":"Garzoni","year":"2009","journal-title":"Exp. Cell Res."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1455","DOI":"10.1590\/S0100-879X2005001000002","article-title":"Multicellular spheroids of bone marrow stromal cells: A three-dimensional in vitro culture system for the study of hematopoietic cell migration","volume":"38","author":"Rossi","year":"2005","journal-title":"Braz. J. Med. Biol. Res."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"260","DOI":"10.1016\/j.yexcr.2017.11.026","article-title":"Inhibition of TGF-\u03b2 pathway reverts extracellular matrix remodeling in T. cruzi-infected cardiac spheroids","volume":"362","author":"Nisimura","year":"2018","journal-title":"Exp. Cell Res."},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Nisimura, L.M., Ferr\u00e3o, P.M., Nogueira, A.D.R., Waghabi, M.C., Meuser-Batista, M., Moreira, O.C., Urbina, J.A., and Garzoni, L.R. (2020). Effect of Posaconazole in an in vitro model of cardiac fibrosis induced by Trypanosoma cruzi. Mol. Biochem. Parasitol., 238.","DOI":"10.1016\/j.molbiopara.2020.111283"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"473","DOI":"10.1016\/j.suc.2012.12.002","article-title":"Adjuvant systemic therapies in breast cancer","volume":"93","year":"2013","journal-title":"Surg. Clin. N. Am."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1483","DOI":"10.1007\/s13346-018-0551-3","article-title":"Metastatic and triple-negative breast cancer: Challenges and treatment options","volume":"8","author":"Sapiezynski","year":"2018","journal-title":"Drug Deliv. Transl. Res."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"263","DOI":"10.1007\/s10585-021-10144-5","article-title":"Current challenges in metastasis research future innovation for clinical translation","volume":"39","author":"Parker","year":"2022","journal-title":"Clin. Exp. Metastasis"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"38","DOI":"10.1186\/s40880-017-0206-7","article-title":"Cancer metastasis: Issues and challenges","volume":"36","author":"Qian","year":"2017","journal-title":"Chin. J. Cancer"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1038\/nrc.2016.25","article-title":"Targeting metastasis","volume":"16","author":"Steeg","year":"2016","journal-title":"Nat. Rev. Cancer"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"4466","DOI":"10.1073\/pnas.74.10.4466","article-title":"Hormonal effects on intracellular and secreted casein in cultures of mouse mammary epithelial cells on floating collagen membranes","volume":"74","author":"Emerman","year":"1977","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_55","doi-asserted-by":"crossref","unstructured":"Halfter, K., Ditsch, N., Kolberg, H.C., Fischer, H., Hauzenberger, T., von Koch, F.E., Bauerfeind, I., von Minckwitz, G., Funke, I., and Crispin, A. (2015). Prospective cohort study using the breast cancer spheroid model as a predictor for response to neoadjuvant therapy\u2014The SpheroNEO study. BMC Cancer, 15.","DOI":"10.1186\/s12885-015-1491-7"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1186\/s12967-016-0855-3","article-title":"Testing chemotherapy efficacy in HER2 negative breast cancer using patient-derived spheroids","volume":"14","author":"Halfter","year":"2016","journal-title":"J. Transl. Med."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"228","DOI":"10.1016\/j.actbio.2021.10.031","article-title":"3D bioprinted drug-resistant breast cancer spheroids for quantitative in situ evaluation of drug resistance","volume":"138","author":"Hong","year":"2022","journal-title":"Acta Biomater."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.actbio.2020.02.006","article-title":"Breast cancer models: Engineering the tumor microenvironment","volume":"106","author":"Bahcecioglu","year":"2020","journal-title":"Acta Biomater."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"50","DOI":"10.1186\/s13058-017-0843-4","article-title":"A 3D in vitro model of the human breast duct: A method to unravel myoepithelial-luminal interactions in the progression of breast cancer","volume":"19","author":"Carter","year":"2017","journal-title":"Breast Cancer Res."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1016\/j.pharmthera.2016.03.013","article-title":"Three-dimensional culture systems in cancer research: Focus on tumor spheroid model","volume":"163","author":"Nath","year":"2016","journal-title":"Pharmacol. Ther."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.neo.2014.12.004","article-title":"Spherical cancer models in tumor biology","volume":"17","author":"Weiswald","year":"2015","journal-title":"Neoplasia"},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"5395","DOI":"10.2147\/OTT.S249756","article-title":"Characterization of Triple-Negative Breast Cancer MDA-MB-231 Cell Spheroid Model","volume":"13","author":"Huang","year":"2020","journal-title":"Onco Targets Ther."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1038\/nprot.2007.30","article-title":"In vitro scratch assay: A convenient and inexpensive method for analysis of cell migration in vitro","volume":"2","author":"Liang","year":"2007","journal-title":"Nat. Protoc."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1016\/j.apsb.2015.07.005","article-title":"Cancer metastases: Challenges and opportunities","volume":"5","author":"Guan","year":"2015","journal-title":"Acta Pharm. Sin. B"},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"394","DOI":"10.1186\/s12935-022-02801-w","article-title":"Current methods for studying metastatic potential of tumor cells","volume":"22","author":"Bouchalova","year":"2022","journal-title":"Cancer Cell Int."},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"lnad007","DOI":"10.1093\/lifemedi\/lnad007","article-title":"Advances in human organoids-on-chips in biomedical research","volume":"2","author":"Wang","year":"2023","journal-title":"Life Med."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"273","DOI":"10.1177\/1087057104265040","article-title":"The use of 3-D cultures for high-throughput screening: The multicellular spheroid model","volume":"9","author":"Freyer","year":"2004","journal-title":"J. Biomol. Screen."},{"key":"ref_68","first-page":"6999","article-title":"Three-dimensional cell culture: A powerful tool in tumor research and drug discovery","volume":"14","author":"Lv","year":"2017","journal-title":"Oncol. Lett."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"1494","DOI":"10.1093\/jnci\/94.19.1494","article-title":"Phenotypic reversion or death of cancer cells by altering signaling pathways in three-dimensional contexts","volume":"94","author":"Wang","year":"2002","journal-title":"J. Natl. Cancer Inst."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"1596","DOI":"10.1242\/jcs.03439","article-title":"The control of tissue architecture over nuclear organization is crucial for epithelial cell fate","volume":"120","author":"Chandramouly","year":"2007","journal-title":"J. Cell Sci."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"1573","DOI":"10.1101\/gad.300566.117","article-title":"Progressive polarity loss and luminal collapse disrupt tissue organization in carcinoma","volume":"31","author":"Halaoui","year":"2017","journal-title":"Genes. Dev."},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"19950","DOI":"10.1038\/s41598-019-56276-3","article-title":"Up-regulated fibronectin in 3D culture facilitates spreading of triple negative breast cancer cells on 2D through integrin \u03b2-5 and Src","volume":"9","author":"Park","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"1383","DOI":"10.1083\/jcb.115.5.1383","article-title":"Control of mammary epithelial differentiation: Basement membrane induces tissue-specific gene expression in the absence of cell-cell interaction and morphological polarity","volume":"115","author":"Streuli","year":"1991","journal-title":"J. Cell Biol."},{"key":"ref_74","doi-asserted-by":"crossref","unstructured":"Correa de Sampaio, P., Auslaender, D., Krubasik, D., Failla, A.V., Skepper, J.N., Murphy, G., and English, W.R. (2012). A heterogeneous in vitro three dimensional model of tumour-stroma interactions regulating sprouting angiogenesis. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0030753"},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1186\/bcr579","article-title":"Polarity determination in breast tissue: Desmosomal adhesion, myoepithelial cells, and laminin 1","volume":"5","author":"Bissell","year":"2003","journal-title":"Breast Cancer Res."},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1046\/j.1365-2613.1998.00051.x","article-title":"Multicellular spheroids: A three-dimensional in vitro culture system to study tumour biology","volume":"79","author":"Kreutz","year":"1998","journal-title":"Int. J. Exp. Pathol."},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"460","DOI":"10.1002\/jcp.10466","article-title":"Three novel hormone-responsive cell lines derived from primary human breast carcinomas: Functional characterization","volume":"199","author":"Mladovan","year":"2004","journal-title":"J. Cell Physiol."},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"1837","DOI":"10.3892\/or.2015.3767","article-title":"Comparison of 2D- and 3D-culture models as drug-testing platforms in breast cancer","volume":"33","author":"Imamura","year":"2015","journal-title":"Oncol. Rep."},{"key":"ref_79","doi-asserted-by":"crossref","unstructured":"Halfter, K., and Mayer, B. (2017). Bringing 3D tumor models to the clinic\u2014Predictive value for personalized medicine. Biotechnol. J., 12.","DOI":"10.1002\/biot.201600295"},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"1420","DOI":"10.1172\/JCI39104","article-title":"The basics of epithelial-mesenchymal transition","volume":"119","author":"Kalluri","year":"2009","journal-title":"J. Clin. Investig."},{"key":"ref_81","doi-asserted-by":"crossref","first-page":"361","DOI":"10.1007\/s11684-018-0656-6","article-title":"Epithelial-to-mesenchymal transition in cancer: Complexity and opportunities","volume":"12","author":"Zhang","year":"2018","journal-title":"Front. Med."},{"key":"ref_82","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1111\/j.2042-7158.2012.01567.x","article-title":"Doxorubicin: An update on anticancer molecular action, toxicity and novel drug delivery systems","volume":"65","author":"Tacar","year":"2013","journal-title":"J. Pharm. Pharmacol."},{"key":"ref_83","doi-asserted-by":"crossref","first-page":"45745","DOI":"10.18632\/oncotarget.9935","article-title":"The relevance of using 3D cell cultures, in addition to 2D monolayer cultures, when evaluating breast cancer drug sensitivity and resistance","volume":"7","author":"Breslin","year":"2016","journal-title":"Oncotarget"},{"key":"ref_84","doi-asserted-by":"crossref","first-page":"951","DOI":"10.1007\/s00432-015-1950-1","article-title":"Evaluation of chemotherapeutics in a three-dimensional breast cancer model","volume":"141","author":"Lovitt","year":"2015","journal-title":"J. Cancer Res. Clin. Oncol."},{"key":"ref_85","doi-asserted-by":"crossref","first-page":"1256","DOI":"10.1016\/j.biotechadv.2014.07.009","article-title":"Three-dimensional in vitro tumor models for cancer research and drug evaluation","volume":"32","author":"Xu","year":"2014","journal-title":"Biotechnol. Adv."},{"key":"ref_86","first-page":"179","article-title":"Development of three-dimensional breast cancer cell culture drug resistance model","volume":"68","author":"Xu","year":"2016","journal-title":"Sheng Li Xue Bao"},{"key":"ref_87","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1007\/s10616-016-0062-3","article-title":"Cytotoxic responses of carnosic acid and doxorubicin on breast cancer cells in butterfly-shaped microchips in comparison to 2D and 3D culture","volume":"69","author":"Anil","year":"2017","journal-title":"Cytotechnology"},{"key":"ref_88","doi-asserted-by":"crossref","unstructured":"Lovitt, C.J., Shelper, T.B., and Avery, V.M. (2018). Doxorubicin resistance in breast cancer cells is mediated by extracellular matrix proteins. BMC Cancer, 18.","DOI":"10.1186\/s12885-017-3953-6"},{"key":"ref_89","doi-asserted-by":"crossref","first-page":"206","DOI":"10.1002\/bit.26845","article-title":"3D tumor spheroids as in vitro models to mimic in vivo human solid tumors resistance to therapeutic drugs","volume":"116","author":"Nunes","year":"2019","journal-title":"Biotechnol. Bioeng."},{"key":"ref_90","first-page":"829","article-title":"Comparison of mammosphere formation from breast cancer cell lines and primary breast tumors","volume":"6","author":"Wang","year":"2014","journal-title":"J. Thorac. Dis."},{"key":"ref_91","doi-asserted-by":"crossref","unstructured":"Yilmazer, A. (2018). Evaluation of cancer stemness in breast cancer and glioblastoma spheroids in vitro. 3 Biotech, 8.","DOI":"10.1007\/s13205-018-1412-y"},{"key":"ref_92","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1159\/000265166","article-title":"Doxorubicin cardiomyopathy","volume":"115","author":"Chatterjee","year":"2010","journal-title":"Cardiology"},{"key":"ref_93","doi-asserted-by":"crossref","first-page":"46663","DOI":"10.18632\/oncotarget.16944","article-title":"Autophagy and mitophagy in the context of doxorubicin-induced cardiotoxicity","volume":"8","author":"Koleini","year":"2017","journal-title":"Oncotarget"},{"key":"ref_94","doi-asserted-by":"crossref","first-page":"R589","DOI":"10.1186\/bcr1259","article-title":"Differential responses to doxorubicin-induced phosphorylation and activation of Akt in human breast cancer cells","volume":"7","author":"Li","year":"2005","journal-title":"Breast Cancer Res."},{"key":"ref_95","doi-asserted-by":"crossref","first-page":"2115","DOI":"10.1158\/1535-7163.MCT-06-0190","article-title":"The analysis of doxorubicin resistance in human breast cancer cells using antibody microarrays","volume":"5","author":"Smith","year":"2006","journal-title":"Mol. Cancer Ther."},{"key":"ref_96","doi-asserted-by":"crossref","first-page":"853","DOI":"10.4103\/0973-1482.139267","article-title":"An overview of doxorubicin formulations in cancer therapy","volume":"10","author":"Rivankar","year":"2014","journal-title":"J. Cancer Res. Ther."},{"key":"ref_97","doi-asserted-by":"crossref","first-page":"1015","DOI":"10.2147\/OTT.S60114","article-title":"Chemotherapy-enhanced inflammation may lead to the failure of therapy and metastasis","volume":"12","author":"Vyas","year":"2014","journal-title":"Onco Targets Ther."},{"key":"ref_98","doi-asserted-by":"crossref","first-page":"2016","DOI":"10.3892\/or.2017.5435","article-title":"Combined effects of furanodiene and doxorubicin on the migration and invasion of MDA-MB-231 breast cancer cells in vitro","volume":"37","author":"Zhong","year":"2017","journal-title":"Oncol. Rep."},{"key":"ref_99","doi-asserted-by":"crossref","unstructured":"Bandyopadhyay, A., Wang, L., Agyin, J., Tang, Y., Lin, S., Yeh, I.T., De, K., and Sun, L.Z. (2010). Doxorubicin in combination with a small TGFbeta inhibitor: A potential novel therapy for metastatic breast cancer in mouse models. PLoS ONE, 5.","DOI":"10.1371\/journal.pone.0010365"},{"key":"ref_100","doi-asserted-by":"crossref","first-page":"185","DOI":"10.4048\/jbc.2019.22.e22","article-title":"Doxorubicin Promotes Migration and Invasion of Breast Cancer Cells through the Upregulation of the RhoA\/MLC Pathway","volume":"22","author":"Liu","year":"2019","journal-title":"J. Breast Cancer"},{"key":"ref_101","doi-asserted-by":"crossref","first-page":"2657","DOI":"10.1158\/1078-0432.CCR-08-2372","article-title":"Twist1-mediated adriamycin-induced epithelial-mesenchymal transition relates to multidrug resistance and invasive potential in breast cancer cells","volume":"15","author":"Li","year":"2009","journal-title":"Clin. Cancer Res."},{"key":"ref_102","doi-asserted-by":"crossref","first-page":"2836","DOI":"10.1111\/febs.13767","article-title":"Low doses of paclitaxel enhance liver metastasis of breast cancer cells in the mouse model","volume":"283","author":"Li","year":"2016","journal-title":"FEBS J."}],"container-title":["Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-7737\/13\/7\/463\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T15:02:41Z","timestamp":1760108561000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-7737\/13\/7\/463"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,6,21]]},"references-count":102,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2024,7]]}},"alternative-id":["biology13070463"],"URL":"https:\/\/doi.org\/10.3390\/biology13070463","relation":{},"ISSN":["2079-7737"],"issn-type":[{"value":"2079-7737","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,6,21]]}}}