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Although species of the\n                    <jats:italic>Ageratum<\/jats:italic>\n                    genus exhibit biological activity, the antitumor potential of\n                    <jats:italic>Ageratum fastigiatum<\/jats:italic>\n                    and its evaluation in three-dimensional (3D) microenvironment have not been previously investigated. This study evaluated the cytotoxic, antiproliferative, and antimigratory effects of the ethanolic extract of\n                    <jats:italic>Ageratum. fastigiatum<\/jats:italic>\n                    on TNBC cells MDA-MB-231 using conventional two-dimensional (2D) cultures and a 3D tumor-on-a-chip model. In 2D cultures, cell viability assessed by the MTT assay revealed an IC\n                    <jats:sub>50<\/jats:sub>\n                    of 308\u00a0\u00b5g\/mL, and approximately 80% reduction in viability at a concentration of 500\u00a0\u00b5g\/mL. The extract significantly inhibited cell migration in scratch assays and reduced colony formation in clonogenic assays. Cell cycle analysis showed an increased sub-G0\/G1 population at the IC\n                    <jats:sub>50<\/jats:sub>\n                    concentration, indicating apoptosis-associated cell cycle disruption. In the 3D microfluidic model, tumor cells embedded in a collagen hydrogel exhibited reduced sensitivity to the extract compared to 2D cultures, with viable cell areas of approximately 15%, compared to 20% under control group , highlighting the influence of the extracellular matrix on treatment response. Additionally, the extract inhibited cell migration by approximately 59% in the 3D model at the IC\n                    <jats:sub>25<\/jats:sub>\n                    concentration (289\u00a0\u00b5g\/mL). In conclusion, the ethanolic extract of\n                    <jats:italic>A. fastigiatum<\/jats:italic>\n                    demonstrated antitumor activity against TNBC cells, and the incorporation of 3D microfluidic models highlights the importance of tumor microenvironment based platforms for a more physiologically relevant evaluation of therapeutic candidates.\n                  <\/jats:p>","DOI":"10.1007\/s12010-026-05589-x","type":"journal-article","created":{"date-parts":[[2026,2,7]],"date-time":"2026-02-07T06:58:13Z","timestamp":1770447493000},"page":"3061-3080","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Effect of Ageratum fastigiatum on Viability, Migration and Proliferation of Breast Cancer Cells in 2D and 3D Culture Models"],"prefix":"10.1007","volume":"198","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-8344-2908","authenticated-orcid":false,"given":"Jo\u00e3o Paulo de Jesus","family":"Vieira","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7493-5376","authenticated-orcid":false,"given":"Ilva de F\u00e1tima","family":"Souza","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8490-7408","authenticated-orcid":false,"given":"Marcelo Br\u00e1ulio","family":"Pedras","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7766-5719","authenticated-orcid":false,"given":"Elton Di\u00eago","family":"Bonif\u00e1cio","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-2321-0075","authenticated-orcid":false,"given":"Michaelle Geralda","family":"dos Santos","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3874-4799","authenticated-orcid":false,"given":"Beth\u00e2nia Alves de","family":"Avelar Freitas","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3355-677X","authenticated-orcid":false,"given":"Libardo Andr\u00e9s","family":"Gonz\u00e1lez Torres","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,2,7]]},"reference":[{"issue":"11","key":"5589_CR1","doi-asserted-by":"publisher","first-page":"151","DOI":"10.2147\/bctt.s176070","volume":"11","author":"Z Momenimovahed","year":"2019","unstructured":"Momenimovahed, Z., & Salehiniya, H. 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