o:37511 Alchemilla vulgaris L. inhibits EMT and suppresses migration and invasion in triple-negative breast cancer en Triple-negative breast cancer (TNBC) is a highly aggressive and extremely invasive type of tumor. To prevent poor prognoses and increase a survival rate, it is important to find an anticancer therapeutic or natural nutritional supplement that prevents and decreases its invasive metastatic potential. The aim of the study was to evaluate Alchemilla vulgaris L. anticancer activity on MDA MB-231 and MDA-MB-468 cells in relation to control normal MRC-5 cell line, focusing on its anti-invasive potential, as there is no data on this issue in triple-negative breast cancer. Phytochemical characterization of aerial parts of the plant was performed by HPLC-PDA analysis and total phenolic and flavonoid compounds determination. The cytotoxic activity was tested with MTT assay, thus modulation of redox status in the cell (O2, GSH) with colorimetric methods. Antimigratory effect was evaluated using Wound healing assay. The expression of genes involved in cancer invasion and epithelial-mesenchymal transition (EMT) were determined at mRNA (SNAI1, VIM, MMP2, MMP9, CYP1B1) and protein (MMP9) level with qPCR method and immunocytochemistry, respectively. Immunocytochemistry was also used in testing BRCA1 and H2A protein expression. The phytochemical analyses showed that the most abundant polyphenolic compound was rutin (4.49 ± 0.06 mg/g), with notable amounts of catechin hydrate (3.59 ± 1.07 mg/g), quercetin-3-O-glucopyranoside (3.21 ± 0.13 mg/g) and ferulic acid (2.73 ± 0.02 mg/g). Results of cytotoxicity showed that plant extract was not significantly cytotoxic on TNBC cells but increased O2 and GSH concentration on dose- and time-dependent manner in all tested cell lines. Furthermore, treatment had strong antimigratory activity as it decreased MDA-MB-231 cell migration by more than 30%, but 15 % on MDA-MB-468 cells. The gene and protein expression of MMP-9 as an indicator of tumor invasion, metastasis, and angiogenesis was significantly suppressed in both TNBC cell lines. Relative gene expression of EMT markers was also significantly suppressed under the treatment. Protein expression of H2A was decreased in both treated cancer cell lines. According to our results Alchemilla vulgaris L. has a strong antimigratory and anti-invasive potential on extremely aggressive TNBC. Treatment showed a selective way of acting, as it demonstrated cancer cells-specific activity in comparation with normal lung fibroblasts. Alchemilla vulgaris L.; triple-negative breast cancer; antimigratory potential; anti invasive potential; EMT 1552099 10.1016/j.sajb.2025.12.017 2026-01-21T09:59:14.412Z 44 yes 46 Aleksandra Nikezić University of Kragujevac, Faculty of Science, Department of Biology and Ecology, Radoja Domanovića 12, 34000 Kragujevac, Serbia Jovana Jovankić University of Kragujevac, Faculty of Science, Department of Biology and Ecology, Radoja Domanovića 12, 34000 Kragujevac, Serbia person Danijela Nikodijevć University of Kragujevac, Faculty of Science, Department of Biology and Ecology, Radoja Domanovića 12, 34000 Kragujevac, Serbia person Milena Milutinović University of Kragujevac, Faculty of Science, Department of Biology and Ecology, Radoja Domanovića 12, 34000 Kragujevac, Serbia person Zorana Dobrijević University of Belgrade, Institute for the Application of Nuclear Energy INEP, Banatska 31b, 11080 Belgrade, Serbia doktor bioloških nauka person 0000-0002-4652-7719 Filip Grbović University of Kragujevac, Faculty of Science, Department of Biology and Ecology, Radoja Domanovića 12, 34000 Kragujevac, Serbia person Nenad Vuković University of Kragujevac, Faculty of Science, Department of Chemistry, Radoja Domanovića 12, 34000 Kragujevac, Serbia person Milena Vukić University of Kragujevac, Faculty of Science, Department of Chemistry, Radoja Domanovića 12, 34000 Kragujevac, Serbia person Danijela Cvetković University of Kragujevac, Faculty of Medical Sciences, Svetozara Markovica 69, 34000 Kragujevac, Serbia person application/pdf 1766471 https://phaidrabg.bg.ac.rs/o:37511 no yes 18 70 92000001 11A29 11A2905 Alchemilla vulgaris L. inhibits EMT and suppresses migration and invasion in triple-negative breast cancer 189 427 441 South African Journal of Botany Holanadija Elsevier B.V. 2026