{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,29]],"date-time":"2026-01-29T23:13:25Z","timestamp":1769728405891,"version":"3.49.0"},"reference-count":96,"publisher":"MDPI AG","issue":"14","license":[{"start":{"date-parts":[[2021,7,15]],"date-time":"2021-07-15T00:00:00Z","timestamp":1626307200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["UIDB\/04423\/2020 and UIDP\/04423\/2020"],"award-info":[{"award-number":["UIDB\/04423\/2020 and UIDP\/04423\/2020"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Fucoxanthin (Fx) is a carotenoid derived from marine organisms that exhibits anticancer activities. However, its role as a potential drug adjuvant in breast cancer (BC) treatment is still poorly explored. Firstly, this study investigated the cytotoxic effects of Fx alone and combined with doxorubicin (Dox) and cisplatin (Cis) on a panel of 2D-cultured BC cell lines (MCF7, SKBR3 and MDA-MB-231) and one non-tumoral cell line (MCF12A). Fucoxanthin induced cytotoxicity against all the cell lines and potentiated Dox cytotoxic effects towards the SKBR3 and MDA-MB-231 cells. The combination triggering the highest cytotoxicity (Fx 10 \u00b5M + Dox 1 \u00b5M in MDA-MB-231) additionally showed significant induction of cell death and genotoxic effects, relative to control. In sequence, the same combination was tested on 3D cultures using a multi-endpoint approach involving bioactivity assays and microscopy techniques. Similar to 2D cultures, the combination of Fx and Dox showed higher cytotoxic effects on 3D cultures compared to the isolated compounds. Furthermore, this combination increased the number of apoptotic cells, decreased cell proliferation, and caused structural and ultrastructural damages on the 3D models. Overall, our findings suggest Fx has potential to become an adjuvant for Dox chemotherapy regimens in BC treatment.<\/jats:p>","DOI":"10.3390\/molecules26144288","type":"journal-article","created":{"date-parts":[[2021,7,15]],"date-time":"2021-07-15T09:32:07Z","timestamp":1626341527000},"page":"4288","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":19,"title":["Fucoxanthin Holds Potential to Become a Drug Adjuvant in Breast Cancer Treatment: Evidence from 2D and 3D Cell Cultures"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4978-5358","authenticated-orcid":false,"given":"Fernanda","family":"Malh\u00e3o","sequence":"first","affiliation":[{"name":"Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto (U.Porto), Rua de Jorge Viterbo Ferreira 228, 4050-313 Porto, Portugal"},{"name":"Interdisciplinary Center for Marine and Environmental Research (CIIMAR), University of Porto (U.Porto), Avenida General Norton de Matos, 4450-208 Matosinhos, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3533-7343","authenticated-orcid":false,"given":"Ana Catarina","family":"Macedo","sequence":"additional","affiliation":[{"name":"Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto (U.Porto), Rua de Jorge Viterbo Ferreira 228, 4050-313 Porto, Portugal"},{"name":"Interdisciplinary Center for Marine and Environmental Research (CIIMAR), University of Porto (U.Porto), Avenida General Norton de Matos, 4450-208 Matosinhos, Portugal"}]},{"given":"Carla","family":"Costa","sequence":"additional","affiliation":[{"name":"Environmental Health Department, National Health Institute Dr. Ricardo Jorge, Rua Alexandre Herculano 321, 4000-055 Porto, Portugal"},{"name":"EPIUnit\u2014Instituto de Sa\u00fade P\u00fablica, University of Porto (U.Porto), Rua das Taipas 135, 4050-600 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9813-4389","authenticated-orcid":false,"given":"Eduardo","family":"Rocha","sequence":"additional","affiliation":[{"name":"Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto (U.Porto), Rua de Jorge Viterbo Ferreira 228, 4050-313 Porto, Portugal"},{"name":"Interdisciplinary Center for Marine and Environmental Research (CIIMAR), University of Porto (U.Porto), Avenida General Norton de Matos, 4450-208 Matosinhos, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2095-359X","authenticated-orcid":false,"given":"Alice Abreu","family":"Ramos","sequence":"additional","affiliation":[{"name":"Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto (U.Porto), Rua de Jorge Viterbo Ferreira 228, 4050-313 Porto, Portugal"},{"name":"Interdisciplinary Center for Marine and Environmental Research (CIIMAR), University of Porto (U.Porto), Avenida General Norton de Matos, 4450-208 Matosinhos, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2021,7,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"629","DOI":"10.1021\/acs.jnatprod.5b01055","article-title":"Natural products as sources of new drugs from 1981 to 2014","volume":"79","author":"Newman","year":"2016","journal-title":"J. Nat. Prod."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1111\/1751-7915.13351","article-title":"Unlocking the potential of natural products in drug discovery","volume":"12","author":"Wright","year":"2019","journal-title":"Microb. Biotechnol."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Sharifi-Rad, J., Ozleyen, A., Boyunegmez Tumer, T., Oluwaseun Adetunji, C., El Omari, N., Balahbib, A., Taheri, Y., Bouyahya, A., Martorell, M., and Martins, N. (2019). Natural products and synthetic analogs as a source of antitumor drugs. Biomolecules, 9.","DOI":"10.3390\/biom9110679"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"354","DOI":"10.2174\/1871520617666171106130325","article-title":"A review of the components of seaweeds as potential candidates in cancer therapy","volume":"18","author":"Ruan","year":"2018","journal-title":"Curr. Med. Chem. Anticancer Agents"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"531","DOI":"10.1016\/j.biotechadv.2011.02.002","article-title":"Gold from the sea: Marine compounds as inhibitors of the hallmarks of cancer","volume":"29","author":"Schumacher","year":"2011","journal-title":"Biotechnol. Adv."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"5","DOI":"10.3390\/md101005","article-title":"Drugs from the sea\u2014Opportunities and obstacles","volume":"1","author":"Proksch","year":"2003","journal-title":"Mar. Drugs"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Khalifa, S.A.M., Elias, N., Farag, M.A., Chen, L., Saeed, A., Hegazy, M.F., Moustafa, M.S., Abd El-Wahed, A., Al-Mousawi, S.M., and Musharraf, S.G. (2019). Marine natural products: A source of novel anticancer drugs. Mar. Drugs, 17.","DOI":"10.3390\/md17090491"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1111\/bph.14876","article-title":"Enriching cancer pharmacology with drugs of marine origin","volume":"177","author":"Jimenez","year":"2020","journal-title":"Br. J. Pharmacol."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Dyshlovoy, S.A., and Honecker, F. (2019). Marine compounds and cancer: The first two decades of XXI century. Mar. Drugs, 18.","DOI":"10.3390\/md18010020"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Maiti, S. (2018). Therapeutic potential of seaweed bioactive compounds. Seaweed Biomaterials, IntechOpen.","DOI":"10.5772\/intechopen.71251"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"771","DOI":"10.1007\/s10811-012-9931-0","article-title":"The consumption of seaweed as a protective factor in the etiology of breast cancer: Proof of principle","volume":"25","author":"Teas","year":"2013","journal-title":"J. Appl. Phycol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1345","DOI":"10.1017\/S0007114509993242","article-title":"A case\u2013control study on seaweed consumption and the risk of breast cancer","volume":"103","author":"Yang","year":"2010","journal-title":"Br. J. Nutr."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"483","DOI":"10.1111\/j.1349-7006.2001.tb01119.x","article-title":"Seaweed prevents breast cancer?","volume":"92","author":"Funahashi","year":"2001","journal-title":"Jpn. J. Cancer Res."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1007\/s10565-014-9277-2","article-title":"Inhibitory effects of fucoxanthinol on the viability of human breast cancer cell lines MCF-7 and MDA-MB-231 are correlated with modulation of the NF-kappaB pathway","volume":"30","author":"Rwigemera","year":"2014","journal-title":"Cell Biol. Toxicol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"158618","DOI":"10.1016\/j.bbalip.2020.158618","article-title":"Health benefits of fucoxanthin in the prevention of chronic diseases","volume":"1865","author":"Bae","year":"2020","journal-title":"Biochim. Biophys. Acta Mol. Cell Biol. Lipids"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"4784","DOI":"10.3390\/md13084784","article-title":"Fucoxanthin and its metabolite fucoxanthinol in cancer prevention and treatment","volume":"13","author":"Martin","year":"2015","journal-title":"Mar. Drugs"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"5130","DOI":"10.3390\/md11125130","article-title":"Fucoxanthin: A marine carotenoid exerting anti-cancer effects by affecting multiple mechanisms","volume":"11","author":"Kumar","year":"2013","journal-title":"Mar. Drugs"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Meresse, S., Fodil, M., Fleury, F., and Chenais, B. (2020). Fucoxanthin, a marine-derived carotenoid from brown seaweeds and microalgae: A promising bioactive compound for cancer therapy. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21239273"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Garg, S., Afzal, S., Elwakeel, A., Sharma, D., Radhakrishnan, N., Dhanjal, J.K., Sundar, D., Kaul, S.C., and Wadhwa, R. (2019). Marine carotenoid fucoxanthin possesses anti-metastasis activity: Molecular evidence. Mar. Drugs, 17.","DOI":"10.3390\/md17060338"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"723515","DOI":"10.1155\/2015\/723515","article-title":"Fucoxanthin: A promising medicinal and nutritional ingredient","volume":"2015","author":"Zhang","year":"2015","journal-title":"Evid. Based Complement. Alternat. Med."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2702","DOI":"10.1002\/ijc.23860","article-title":"Anti-adult T-cell leukemia effects of brown algae fucoxanthin and its deacetylated product, fucoxanthinol","volume":"123","author":"Ishikawa","year":"2008","journal-title":"Int. J. Cancer"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1557","DOI":"10.21873\/anticanres.11484","article-title":"Antitumor and cancer-preventative function of fucoxanthin: A marine carotenoid","volume":"37","author":"Satomi","year":"2017","journal-title":"Anticancer Res."},{"key":"ref_23","first-page":"207","article-title":"Comparative effects between fucoxanthinol and its precursor fucoxanthin on viability and apoptosis of breast cancer cell lines MCF-7 and MDA-MB-231","volume":"35","author":"Rwigemera","year":"2015","journal-title":"Anticancer Res."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"45","DOI":"10.5493\/wjem.v2.i3.45","article-title":"Natural compounds as anticancer agents: Experimental evidence","volume":"2","author":"Wang","year":"2012","journal-title":"World J. Exp. Med."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2219","DOI":"10.1111\/jcmm.14151","article-title":"Fucoxanthin inhibits tumour-related lymphangiogenesis and growth of breast cancer","volume":"23","author":"Wang","year":"2019","journal-title":"J. Cell Mol. Med."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Sung, H., Ferlay, J., Siegel, R.L., Laversanne, M., Soerjomataram, I., Jemal, A., and Bray, F. (2021). Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J. Clin.","DOI":"10.3322\/caac.21660"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"7","DOI":"10.3322\/caac.21590","article-title":"Cancer statistics, 2020","volume":"70","author":"Siegel","year":"2020","journal-title":"CA Cancer J. Clin."},{"key":"ref_28","unstructured":"World Health Organization (2021, May 01). Breast Cancer\u2014WHO|World Health Organization. Available online: https:\/\/www.who.int\/news-room\/fact-sheets\/detail\/breast-cancer."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1016\/j.soc.2017.08.005","article-title":"Molecular Subtypes and Local-Regional Control of Breast Cancer","volume":"27","author":"Fragomeni","year":"2018","journal-title":"Surg. Oncol. Clin. N. Am."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2206","DOI":"10.1093\/annonc\/mdt303","article-title":"Personalizing the treatment of women with early breast cancer: Highlights of the St Gallen International Expert Consensus on the primary therapy of early breast cancer 2013","volume":"24","author":"Goldhirsch","year":"2013","journal-title":"Ann. Oncol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"412","DOI":"10.5306\/wjco.v5.i3.412","article-title":"Biological subtypes of breast cancer: Prognostic and therapeutic implications","volume":"5","author":"Yersal","year":"2014","journal-title":"World J. Clin. Oncol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"172","DOI":"10.1016\/j.breast.2007.08.009","article-title":"Roles of hormone replacement therapy and iron in proliferation of breast epithelial cells with different estrogen and progesterone receptor status","volume":"17","author":"Dai","year":"2008","journal-title":"Breast"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1993","DOI":"10.1007\/s40265-015-0489-4","article-title":"Recent advances in the development of antineoplastic agents derived from natural products","volume":"75","author":"Trendowski","year":"2015","journal-title":"Drugs"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"176","DOI":"10.1158\/2159-8290.CD-18-1177","article-title":"Insights into molecular classifications of triple-negative breast cancer: Improving patient selection for treatment","volume":"9","author":"Lin","year":"2019","journal-title":"Cancer Discov."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1186\/s13058-020-01296-5","article-title":"Triple-negative breast cancer molecular subtyping and treatment progress","volume":"22","author":"Yin","year":"2020","journal-title":"Breast Cancer Res."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"3","DOI":"10.2174\/2211738507666190122111224","article-title":"Drug combinations in breast cancer therapy","volume":"7","author":"Fisusi","year":"2019","journal-title":"Pharm. Nanotechnol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"725","DOI":"10.1093\/carcin\/bgt086","article-title":"A tale of two approaches: Complementary mechanisms of cytotoxic and targeted therapy resistance may inform next-generation cancer treatments","volume":"34","author":"Masui","year":"2013","journal-title":"Carcinogenesis"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1016\/j.phrs.2016.01.001","article-title":"Cisplatin resistance and opportunities for precision medicine","volume":"106","author":"Amable","year":"2016","journal-title":"Pharmacol. Res."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1016\/j.phytol.2015.09.007","article-title":"Bioactive compounds from brown seaweeds: Phloroglucinol, fucoxanthin and fucoidan as promising therapeutic agents against breast cancer","volume":"14","author":"Rocha","year":"2015","journal-title":"PhytoChem. Lett."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"5261","DOI":"10.21873\/anticanres.13721","article-title":"Therapeutic application of diverse marine-derived natural products in cancer therapy","volume":"39","author":"Yun","year":"2019","journal-title":"Anticancer Res."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"776","DOI":"10.1080\/15287394.2017.1357297","article-title":"Anticancer effects of seaweed compounds fucoxanthin and phloroglucinol, alone and in combination with 5-fluorouracil in colon cells","volume":"80","author":"Abreu","year":"2017","journal-title":"J. Toxicol. Environ. Health A"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1016\/j.etap.2018.02.006","article-title":"Cytotoxic activity of fucoxanthin, alone and in combination with the cancer drugs imatinib and doxorubicin, in CML cell lines","volume":"59","author":"Almeida","year":"2018","journal-title":"Environ. Toxicol. Pharmacol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"675","DOI":"10.1016\/j.fct.2018.06.027","article-title":"Low-dose doxorubicin with carotenoids selectively alters redox status and upregulates oxidative stress-mediated apoptosis in breast cancer cells","volume":"118","author":"Vijay","year":"2018","journal-title":"Food Chem. Toxicol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1007\/s00432-019-02841-2","article-title":"The anticancer effects and mechanisms of fucoxanthin combined with other drugs","volume":"145","author":"Wang","year":"2019","journal-title":"J. Cancer Res. Clin. Oncol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1093\/nutrit\/nuy066","article-title":"Risks and benefits of consuming edible seaweeds","volume":"77","author":"Cherry","year":"2019","journal-title":"Nutr. Rev."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1023\/B:BREA.0000025418.88785.2b","article-title":"Three-dimensional in vitro tissue culture models of breast cancer\u2014A review","volume":"85","author":"Kim","year":"2004","journal-title":"Breast Cancer Res. Treat."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"2993","DOI":"10.1002\/jcp.26052","article-title":"Three-dimensional cell culture models for anticancer drug screening: Worth the effort?","volume":"233","author":"Verjans","year":"2018","journal-title":"J. Cell Physiol."},{"key":"ref_48","first-page":"910","article-title":"2D and 3D cell cultures\u2014A comparison of different types of cancer cell cultures","volume":"14","author":"Kolenda","year":"2018","journal-title":"Arch. Med. Sci."},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Santo, V.E., Rebelo, S.P., Estrada, M.F., Alves, P.M., Boghaert, E., and Brito, C. (2017). Drug screening in 3D in vitro tumor models: Overcoming current pitfalls of efficacy read-outs. Biotechnol. J., 12.","DOI":"10.1002\/biot.201600505"},{"key":"ref_50","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_51","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1016\/j.cbpc.2005.10.005","article-title":"Halocynthiaxanthin and fucoxanthinol isolated from Halocynthia roretzi induce apoptosis in human leukemia, breast and colon cancer cells","volume":"142","author":"Konishi","year":"2006","journal-title":"Comp. BioChem. Physiol. C Toxicol. Pharmacol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1016\/j.canlet.2010.10.016","article-title":"Fucoxanthin and its deacetylated product, fucoxanthinol, induce apoptosis of primary effusion lymphomas","volume":"300","author":"Yamamoto","year":"2011","journal-title":"Cancer Lett."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1186\/1475-2867-13-39","article-title":"Assessment of potential anti-cancer stem cell activity of marine algal compounds using an in vitro mammosphere assay","volume":"13","author":"Sterrenberg","year":"2013","journal-title":"Cancer Cell Int."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"336","DOI":"10.1111\/j.1742-7843.2006.pto_167.x","article-title":"The alkaline comet assay: Towards validation in biomonitoring of DNA damaging exposures","volume":"98","year":"2006","journal-title":"Basic. Clin. Pharmacol. Toxicol."},{"key":"ref_55","first-page":"3025","article-title":"Resveratrol decreases Rad51 expression and sensitizes cisplatin-resistant MCF-7 breast cancer cells","volume":"39","author":"Herrera","year":"2018","journal-title":"Oncol. Rep."},{"key":"ref_56","doi-asserted-by":"crossref","unstructured":"Lee, K.S., Lee, M.G., Kwon, Y.S., and Nam, K.S. (2020). Arctigenin enhances the cytotoxic effect of doxorubicin in MDA-MB-231 breast cancer cells. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21082997"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"128","DOI":"10.1016\/j.euprot.2015.05.002","article-title":"Pathway-centric analysis of the DNA damage response to chemotherapeutic agents in two breast cell lines","volume":"8","author":"Antberg","year":"2015","journal-title":"EuPA Open Proteom."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"753","DOI":"10.3892\/ijo.2016.3558","article-title":"Influence of doxorubicin on apoptosis and oxidative stress in breast cancer cell lines","volume":"49","author":"Calaf","year":"2016","journal-title":"Int. J. Oncol."},{"key":"ref_59","first-page":"84","article-title":"Doxorubicin, DNA torsion, and chromatin dynamics","volume":"1845","author":"Yang","year":"2014","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"300","DOI":"10.1159\/000245937","article-title":"Doxorubicin exerts cytotoxic effects through cell cycle arrest and Fas-mediated cell death","volume":"84","author":"Kim","year":"2009","journal-title":"Pharmacology"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"1409","DOI":"10.1111\/bph.14816","article-title":"Natural compounds as potential adjuvants to cancer therapy: Preclinical evidence","volume":"177","author":"Lin","year":"2020","journal-title":"Br. J. Pharmacol."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"3489","DOI":"10.1158\/1078-0432.CCR-16-3083","article-title":"Clinically relevant concentrations of anticancer drugs: A guide for nonclinical studies","volume":"23","author":"Liston","year":"2017","journal-title":"Clin. Cancer Res."},{"key":"ref_63","doi-asserted-by":"crossref","unstructured":"Men\u00e9ndez-Men\u00e9ndez, J., Hermida-Prado, F., Granda-D\u00edaz, R., Gonz\u00e1lez, A., Garc\u00eda-Pedrero, J.M., Del-R\u00edo-Ibisate, N., Gonz\u00e1lez-Gonz\u00e1lez, A., Cos, S., Alonso-Gonz\u00e1lez, C., and Mart\u00ednez-Campa, C. (2019). Deciphering the molecular basis of melatonin protective effects on breast cells treated with doxorubicin: TWIST1 a transcription factor involved in EMT and metastasis, a novel target of melatonin. Cancers, 11.","DOI":"10.3390\/cancers11071011"},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"134","DOI":"10.3389\/fonc.2013.00134","article-title":"Cisplatin induces differentiation of breast cancer cells","volume":"3","author":"Prabhakaran","year":"2013","journal-title":"Front. Oncol."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1016\/j.bbagen.2004.08.012","article-title":"Fucoxanthin induces apoptosis and enhances the antiproliferative effect of the PPARgamma ligand, troglitazone, on colon cancer cells","volume":"1675","author":"Hosokawa","year":"2004","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"877","DOI":"10.1093\/abbs\/gmu080","article-title":"Fucoxanthin induces growth arrest and apoptosis in human bladder cancer T24 cells by up-regulation of p21 and down-regulation of mortalin","volume":"46","author":"Wang","year":"2014","journal-title":"Acta Biochim. Biophys. Sin."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"2212","DOI":"10.1021\/acs.jafc.8b07126","article-title":"Induction of apoptosis in human glioma cells by fucoxanthin via triggering of ROS-mediated oxidative damage and regulation of MAPKs and PI3K-AKT pathways","volume":"67","author":"Wu","year":"2019","journal-title":"J. Agric. Food Chem."},{"key":"ref_68","doi-asserted-by":"crossref","unstructured":"Shin, J., Song, M.-H., Oh, J.-W., Keum, Y.-S., and Saini, R.K. (2020). Pro-oxidant actions of carotenoids in triggering apoptosis of cancer cells: A review of emerging evidence. Antioxidants, 9.","DOI":"10.3390\/antiox9060532"},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"11344","DOI":"10.1021\/jf2029785","article-title":"Fucoxanthin enhances HO-1 and NQO1 expression in murine hepatic BNL CL.2 cells through activation of the Nrf2\/ARE system partially by its pro-oxidant activity","volume":"59","author":"Liu","year":"2011","journal-title":"J. Agric. Food Chem."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"540","DOI":"10.3389\/fbioe.2020.00540","article-title":"Human melanocyte-derived spheroids: A precise test system for drug screening and a multicellular unit for tissue engineering","volume":"8","author":"Zurina","year":"2020","journal-title":"Front. Bioeng. Biotechnol."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1080\/01635581.2017.1339814","article-title":"Induction of anoikis in human colorectal cancer cells by fucoxanthinol","volume":"69","author":"Terasaki","year":"2017","journal-title":"Nutr. Cancer"},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"52","DOI":"10.3164\/jcbn.18-45","article-title":"Fucoxanthin administration delays occurrence of tumors in xenograft mice by colonospheres, with an anti-tumor predictor of glycine","volume":"64","author":"Terasaki","year":"2019","journal-title":"J. Clin. BioChem. Nutr."},{"key":"ref_73","doi-asserted-by":"crossref","unstructured":"Hongisto, V., Jernstr\u00f6m, S., Fey, V., Mpindi, J.P., Kleivi Sahlberg, K., Kallioniemi, O., and Per\u00e4l\u00e4, M. (2013). High-throughput 3D screening reveals differences in drug sensitivities between culture models of JIMT1 breast cancer cells. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0077232"},{"key":"ref_74","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_75","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/978-1-4939-6960-9_1","article-title":"Basic. colorimetric proliferation assays: MTT, WST, and Resazurin","volume":"1601","author":"Prabst","year":"2017","journal-title":"Methods Mol. Biol."},{"key":"ref_76","doi-asserted-by":"crossref","unstructured":"Kumar, P., Nagarajan, A., and Uchil, P.D. (2018). Analysis of cell viability by the lactate dehydrogenase assay. Cold Spring Harb. Protoc., 2018.","DOI":"10.1101\/pdb.prot095497"},{"key":"ref_77","doi-asserted-by":"crossref","unstructured":"Malh\u00e3o, F., Ramos, A.A., Macedo, A.C., and Rocha, E. (2021). Cytotoxicity of seaweed compounds, alone or combined to reference drugs, against breast cell lines cultured in 2D and 3D. Toxics, 9.","DOI":"10.3390\/toxics9020024"},{"key":"ref_78","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":"Oncol. Targets Ther."},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"942","DOI":"10.1038\/sj.cdd.4401556","article-title":"Apoptosis-based therapies and drug targets","volume":"12","author":"Fischer","year":"2005","journal-title":"Cell Death Differ."},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"384","DOI":"10.1007\/s11307-014-0802-8","article-title":"Imaging caspase-3 activation as a marker of apoptosis-targeted treatment response in cancer","volume":"17","author":"Chen","year":"2015","journal-title":"Mol. Imaging Biol."},{"key":"ref_81","doi-asserted-by":"crossref","first-page":"341","DOI":"10.1007\/s10549-017-4270-0","article-title":"Ki67 expression in invasive breast cancer: The use of tissue microarrays compared with whole tissue sections","volume":"164","author":"Muftah","year":"2017","journal-title":"Breast Cancer Res. Treat."},{"key":"ref_82","doi-asserted-by":"crossref","first-page":"496","DOI":"10.20892\/j.issn.2095-3941.2016.0066","article-title":"Ki-67 as a prognostic marker according to breast cancer molecular subtype","volume":"13","author":"Soliman","year":"2016","journal-title":"Cancer Biol. Med."},{"key":"ref_83","doi-asserted-by":"crossref","unstructured":"Hershey, B.J., Vazzana, R., Joppi, D.L., and Havas, K.M. (2019). Lipid droplets define a sub-population of breast cancer stem cells. J. Clin. Med., 9.","DOI":"10.3390\/jcm9010087"},{"key":"ref_84","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1007\/s00418-009-0615-z","article-title":"Quantitative electron microscopy shows uniform incorporation of triglycerides into existing lipid droplets","volume":"132","author":"Cheng","year":"2009","journal-title":"HistoChem. Cell Biol."},{"key":"ref_85","doi-asserted-by":"crossref","first-page":"2196","DOI":"10.3390\/md13042196","article-title":"Anti-obesity activity of the marine carotenoid fucoxanthin","volume":"13","author":"Gammone","year":"2015","journal-title":"Mar. Drugs"},{"key":"ref_86","doi-asserted-by":"crossref","unstructured":"Shyu, P., Wong, X., Fah, A., Crasta, K., and Thibault, G. (2018). Dropping in on lipid droplets: Insights into cellular stress and cancer. Biosci. Rep., 38.","DOI":"10.1042\/BSR20180764"},{"key":"ref_87","doi-asserted-by":"crossref","unstructured":"Bojko, A., Staniak, K., Czarnecka-Herok, J., Sunderland, P., Dudkowska, M., \u015aliwi\u0144ska, M.A., Salmina, K., and Sikora, E. (2020). Improved autophagic flux in escapers from doxorubicin-induced senescence\/polyploidy of breast cancer cells. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21176084"},{"key":"ref_88","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1186\/bcr2889","article-title":"Choosing the right cell line for breast cancer research","volume":"13","author":"Holliday","year":"2011","journal-title":"Breast Cancer Res."},{"key":"ref_89","first-page":"35","article-title":"The expression patterns of ER, PR, HER2, CK5\/6, EGFR, Ki-67 and AR by immunohistochemical analysis in breast cancer cell lines","volume":"4","author":"Subik","year":"2010","journal-title":"Breast Cancer"},{"key":"ref_90","doi-asserted-by":"crossref","unstructured":"Malh\u00e3o, F., Ramos, A.A., Buttachon, S., Dethoup, T., Kijjoa, A., and Rocha, E. (2019). Cytotoxic and antiproliferative effects of preussin, a hydroxypyrrolidine derivative from the marine sponge-associated fungus Aspergillus candidus KUFA 0062, in a panel of breast cancer cell lines and using 2D and 3D cultures. Mar. Drugs, 17.","DOI":"10.3390\/md17080448"},{"key":"ref_91","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1016\/j.cmpb.2015.02.006","article-title":"AnaSP: A software suite for automatic image analysis of multicellular spheroids","volume":"119","author":"Piccinini","year":"2015","journal-title":"Comput. Methods Programs Biomed."},{"key":"ref_92","doi-asserted-by":"crossref","first-page":"747","DOI":"10.1016\/j.tibs.2018.08.004","article-title":"Ki-67: More hidden behind a \u2018classic proliferation marker\u2019","volume":"43","author":"Vagnarelli","year":"2018","journal-title":"Trends BioChem. Sci."},{"key":"ref_93","doi-asserted-by":"crossref","first-page":"289","DOI":"10.1369\/jhc.2008.952044","article-title":"Assessment of apoptosis by immunohistochemistry to active caspase-3, active caspase-7, or cleaved PARP in monolayer cells and spheroid and subcutaneous xenografts of human carcinoma","volume":"57","author":"Bressenot","year":"2009","journal-title":"J. HistoChem. CytoChem."},{"key":"ref_94","first-page":"1","article-title":"PAST: Paleontological Statistics Software Package for education and data analysis","volume":"4","author":"Hammer","year":"2001","journal-title":"Palaeontol. Electron."},{"key":"ref_95","unstructured":"Gaetano, J. (2021, June 01). Holm-Bonferroni Sequential Correction: An Excel Calculator (v. 1.3) Microsoft Excel Workbook. Available online: https:\/\/www.researchgate.net\/publication\/322569220_Holm-Bonferroni_sequential_correction_An_Excel_calculator_13."},{"key":"ref_96","first-page":"65","article-title":"A simple sequentially rejective multiple test procedure","volume":"6","author":"Holm","year":"1979","journal-title":"Scand. Stat. Theory Appl."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/26\/14\/4288\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T06:30:53Z","timestamp":1760164253000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/26\/14\/4288"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,7,15]]},"references-count":96,"journal-issue":{"issue":"14","published-online":{"date-parts":[[2021,7]]}},"alternative-id":["molecules26144288"],"URL":"https:\/\/doi.org\/10.3390\/molecules26144288","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,7,15]]}}}