{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T18:39:54Z","timestamp":1790361594980,"version":"4.1.0"},"reference-count":45,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2026,2,4]],"date-time":"2026-02-04T00:00:00Z","timestamp":1770163200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Andalusian government, FEDER EU and Universidad de M\u00e1laga","award":["PPRO-BIO267-G-2023"],"award-info":[{"award-number":["PPRO-BIO267-G-2023"]}]},{"DOI":"10.13039\/100009473","name":"Universidad de M\u00e1laga","doi-asserted-by":"crossref","award":["PPRO-B4-2024-009"],"award-info":[{"award-number":["PPRO-B4-2024-009"]}],"id":[{"id":"10.13039\/100009473","id-type":"DOI","asserted-by":"crossref"}]},{"name":"A.4. Fellowship"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The tumor microenvironment plays a critical role in cancer progression, with oxidative stress, autophagy, angiogenesis, and cell migration acting as tightly interconnected processes. Natural bioactive compounds have emerged as promising modulators of these pathways; however, their cell type-specific effects within the TME remain poorly understood. In this study, we investigate the effects of punicalin on triple-negative breast cancer and endothelial cells, with a focus on redox homeostasis and autophagy as upstream regulatory mechanisms. Punicalin reduced oxidative stress in MDA-MB-231 cells under basal conditions and strongly attenuated hydrogen peroxide-induced stress, whereas HMEC-1 cells exhibited concentration- and condition-dependent reactive oxygen species (ROS) modulation. Autophagy assays revealed no significant modulation in tumor cells, while a consistent and pronounced decrease in autophagic activity was observed in endothelial cells under both basal and nutrient-deprivation conditions. Functionally, punicalin decreased tumor cell migration and impaired HMEC-1 migration, while HUVEC migration remained largely unaffected. Tube formation assays demonstrated significant inhibition of angiogenic capacity. Taken together, these findings demonstrate that punicalin selectively modulates oxidative stress and autophagy, leading to functional alterations in migration and angiogenesis. By highlighting its selective impact on microvascular endothelial cells while sparing normal endothelium, this study provides a strong rationale for further preclinical evaluation of punicalin.<\/jats:p>","DOI":"10.3390\/ijms27031533","type":"journal-article","created":{"date-parts":[[2026,2,4]],"date-time":"2026-02-04T12:51:56Z","timestamp":1770209516000},"page":"1533","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Punicalin Modulates Angiogenesis and Tumor Microenvironment-Related Processes in Triple-Negative Breast Cancer and Endothelial Cells"],"prefix":"10.3390","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0009-0006-5738-0835","authenticated-orcid":false,"given":"Maria Carmen","family":"Banqueri-Pegalajar","sequence":"first","affiliation":[{"name":"Department of Molecular Biology and Biochemistry, Faculty of Science, Universidad de M\u00e1laga, Andaluc\u00eda Tech, 29071 M\u00e1laga, Spain"},{"name":"Molecular Bases of Biological Systems (SIBIUMA) Group, Instituto de Investigaci\u00f3n Biom\u00e9dica de M\u00e1laga y Plataforma en Nanomedicina-IBIMA Plataforma Bionand, 29590 M\u00e1laga, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-0354-3265","authenticated-orcid":false,"given":"Joel D.","family":"Posligua-Garc\u00eda","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology and Biochemistry, Faculty of Science, Universidad de M\u00e1laga, Andaluc\u00eda Tech, 29071 M\u00e1laga, Spain"},{"name":"Molecular Bases of Biological Systems (SIBIUMA) Group, Instituto de Investigaci\u00f3n Biom\u00e9dica de M\u00e1laga y Plataforma en Nanomedicina-IBIMA Plataforma Bionand, 29590 M\u00e1laga, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-7331-237X","authenticated-orcid":false,"given":"Carlos Ulises","family":"C\u00e1rdenas-Vela","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology and Biochemistry, Faculty of Science, Universidad de M\u00e1laga, Andaluc\u00eda Tech, 29071 M\u00e1laga, Spain"},{"name":"Molecular Bases of Biological Systems (SIBIUMA) Group, Instituto de Investigaci\u00f3n Biom\u00e9dica de M\u00e1laga y Plataforma en Nanomedicina-IBIMA Plataforma Bionand, 29590 M\u00e1laga, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0412-3035","authenticated-orcid":false,"given":"Manuel","family":"Bernal","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology and Biochemistry, Faculty of Science, Universidad de M\u00e1laga, Andaluc\u00eda Tech, 29071 M\u00e1laga, Spain"},{"name":"Molecular Bases of Biological Systems (SIBIUMA) Group, Instituto de Investigaci\u00f3n Biom\u00e9dica de M\u00e1laga y Plataforma en Nanomedicina-IBIMA Plataforma Bionand, 29590 M\u00e1laga, Spain"},{"name":"Center for Biomedical Network Research in Rare Diseases (CIBERER), Instituto de Salud Carlos III, 28029 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7275-6462","authenticated-orcid":false,"given":"Miguel \u00c1ngel","family":"Medina","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology and Biochemistry, Faculty of Science, Universidad de M\u00e1laga, Andaluc\u00eda Tech, 29071 M\u00e1laga, Spain"},{"name":"Molecular Bases of Biological Systems (SIBIUMA) Group, Instituto de Investigaci\u00f3n Biom\u00e9dica de M\u00e1laga y Plataforma en Nanomedicina-IBIMA Plataforma Bionand, 29590 M\u00e1laga, Spain"},{"name":"Center for Biomedical Network Research in Rare Diseases (CIBERER), Instituto de Salud Carlos III, 28029 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,2,4]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"De Cicco, P., Catani, M.V., Gasperi, V., Sibilano, M., Quaglietta, M., and Savini, I. (2019). Nutrition and breast cancer: A literature review on prevention, treatment and recurrence. Nutrients, 11.","DOI":"10.3390\/nu11071514"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Fan, Y., Wang, H., Zhang, H., Na, T., and Zhao, Y. (2025). Integrating molecular targeting and immune modulation in triple-negative breast cancer: From mechanistic insights to therapeutic innovation. Front. Immunol., 16.","DOI":"10.3389\/fimmu.2025.1711415"},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Sarcinella, A., Guerra Villacis, J.S., and Brizzi, M.F. (2025). Fueling the seed: Growth factors and cytokines driving cancer stem cells in gynecological malignancies. Int. J. Mol. Sci., 26.","DOI":"10.3390\/ijms262311462"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1097\/PAP.0000000000000219","article-title":"Paget\u2019s \u201cseed and soil\u201d theory of cancer metastasis: An idea whose time has come","volume":"26","author":"Akhtar","year":"2019","journal-title":"Adv. Anat. Pathol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"374","DOI":"10.1016\/j.ccell.2023.02.016","article-title":"The evolving tumor microenvironment: From cancer initiation to metastatic outgrowth","volume":"41","author":"Joyce","year":"2023","journal-title":"Cancer Cell"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1007\/s12307-009-0025-8","article-title":"The tumor microenvironment: The making of a paradigm","volume":"2","author":"Witz","year":"2009","journal-title":"Cancer Microenviron."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"107753","DOI":"10.1016\/j.pharmthera.2020.107753","article-title":"Tumor microenvironment as a therapeutic target in cancer","volume":"221","author":"Xiao","year":"2021","journal-title":"Pharmacol. Ther."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"108253","DOI":"10.1016\/j.pharmthera.2022.108253","article-title":"Triple negative breast cancer (TNBC): Non-genetic tumor heterogeneity and immune microenvironment: Emerging treatment options","volume":"237","author":"So","year":"2022","journal-title":"Pharmacol. Ther."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"366","DOI":"10.1016\/j.bcp.2019.07.019","article-title":"The strigolactone analog GR-24 inhibits angiogenesis in vivo and in vitro by a mechanism involving cytoskeletal reorganization and VEGFR2 signalling","volume":"168","author":"Carrillo","year":"2019","journal-title":"Biochem. Pharmacol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"665","DOI":"10.1038\/s41418-019-0287-8","article-title":"Autophagy in endothelial cells and tumor angiogenesis","volume":"26","author":"Schaaf","year":"2019","journal-title":"Cell Death Differ."},{"key":"ref_11","first-page":"188352","article-title":"Control of tumor angiogenesis and metastasis through modulation of cell redox state","volume":"1873","author":"Serrano","year":"2020","journal-title":"BBA Rev. Cancer"},{"key":"ref_12","first-page":"189417","article-title":"A primer on metabolic contributions in tumor angiogenic microenvironment","volume":"1880","author":"Bernal","year":"2025","journal-title":"BBA Rev. Cancer"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Boujhoud, Z., Eleroui, M., Feki, A., Saad, H.B., Kraiem, M., Youlyouz Marfak, I., Essayagh, S., Hilali, S., Badraoui, R., and Kallel, H. (2025). Pro-Angiogenic and Wound-Healing Potential of Bioactive Polysaccharides Extracted from Moroccan Algae Osmundea pinnatifida. Life, 15.","DOI":"10.3390\/life15101564"},{"key":"ref_14","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_15","doi-asserted-by":"crossref","first-page":"560","DOI":"10.1038\/s41580-023-00585-z","article-title":"Autophagy and autophagy-related pathways in cancer","volume":"24","author":"Debnath","year":"2023","journal-title":"Nat. Rev. Mol. Cell. Biol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1021","DOI":"10.1111\/1440-1681.12649","article-title":"Autophagy in vascular endothelial cells","volume":"43","author":"Jiang","year":"2016","journal-title":"Clin. Exp. Pharmacol. Physiol."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Serrano, J.J., and Medina, M.A. (2025). Metabolic reprogramming at the edge of redox: Connections between metabolic reprogramming and cancer redox state. Int. J. Mol. Sci., 26.","DOI":"10.3390\/ijms26020498"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"3945","DOI":"10.1111\/cas.15068","article-title":"Reactive oxygen species in cancer: Current findings and future directions","volume":"112","author":"Nakamura","year":"2021","journal-title":"Cancer Sci."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"280","DOI":"10.1038\/s41568-021-00435-0","article-title":"The role of ROS in tumour development and progression","volume":"22","author":"Cheung","year":"2022","journal-title":"Nat. Rev. Cancer"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1603","DOI":"10.1016\/j.freeradbiomed.2010.09.006","article-title":"Oxidative stress, inflammation, and cancer: How are they linked?","volume":"49","author":"Reuter","year":"2010","journal-title":"Free Rad. Biol. Med."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Garc\u00eda-Caballero, M., Torres-Vargas, J.A., Marrero, A.D., Mart\u00ednez-Poveda, B., Medina, M.\u00c1., and Quesada, A.R. (2022). Angioprevention of urologic cancers by plant-derived foods. Pharmaceutics, 14.","DOI":"10.3390\/pharmaceutics14020256"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Marrero, A.D., Quesada, A.R., Mart\u00ednez-Poveda, B., and Medina, M.\u00c1. (2022). Antiangiogenic phytochemicals constituent of diet as promising candidates for chemoprevention of cancer. Antioxidants, 11.","DOI":"10.3390\/antiox11020302"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"694","DOI":"10.2174\/1389201024666230822090318","article-title":"Natural polyphenols in cancer management: Promising role, mechanisms, and chemistry","volume":"25","author":"Upadhyay","year":"2024","journal-title":"Curr. Pharm. Biotechnol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"694","DOI":"10.2174\/1871520621666210726143553","article-title":"An insight into anticancer bioactives from Punica granatum (pomegranate)","volume":"22","author":"Sharma","year":"2022","journal-title":"Anticancer Agents Med. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Li, T., Jiang, G., Hu, X., Yang, D., Tan, T., Gao, Z., Chen, Z., Xiang, C., Li, S., and Ouyang, Z. (2021). Punicalin attenuates breast cancer-associated osteolysis by inhibiting the NF-\u03baB signaling pathway of osteoclasts. Front. Pharmacol., 12, Correction in Front. Pharmacol. 2026, 17, 1678237.","DOI":"10.3389\/fphar.2026.1678237"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"363","DOI":"10.31557\/APJCP.2022.23.1.363","article-title":"Newly synthesized punicalin and punicalagin nano-prototypes induce breast cancer cytotoxicity through ROS-mediated apoptosis","volume":"23","author":"Shalby","year":"2022","journal-title":"Asian Pac. J. Cancer Prev."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"57","DOI":"10.3892\/ijo.2024.5645","article-title":"Beyond self-eating: Emerging autophagy-independent functions for the autophagy molecules in cancer (review)","volume":"64","author":"Tedesco","year":"2024","journal-title":"Int. J. Oncol."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Ma, M.K., and Li, D.Q. (2025). Autophagy-related proteins in triple-negative breast cancer: From molecular insights to therapeutic applications. Int. J. Mol. Sci., 26.","DOI":"10.3390\/ijms26189231"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1497","DOI":"10.1002\/1873-3468.14087","article-title":"Endothelial cell autophagy in homeostasis and cancer","volume":"595","author":"Verhoeven","year":"2021","journal-title":"FEBS Lett."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"176568","DOI":"10.1016\/j.ejphar.2024.176568","article-title":"Novel combinatorial autophagy inhibition therapy for triple negative breast cancers","volume":"973","author":"McKay","year":"2024","journal-title":"Eur. J. Pharmacol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"e18028","DOI":"10.15252\/emmm.202318028","article-title":"Tumor endothelial cell autophagy is a key vascular-immune checkpoint in melanoma","volume":"15","author":"Verhoeven","year":"2023","journal-title":"EMBO Mol. Med."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"789","DOI":"10.1111\/j.2042-7158.1998.tb07141.x","article-title":"Antioxidant and hepatoprotective activity of punicalagin and punicalin on carbon tetrachloride-induced liver damage in rats","volume":"50","author":"Lin","year":"1998","journal-title":"J. Pharm. Pharmacol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"219","DOI":"10.1186\/s12943-025-02375-x","article-title":"Oxidative stress in cancer: From tumor and microenvironment remodeling to therapeutic frontiers","volume":"24","author":"Liang","year":"2025","journal-title":"Mol. Cancer"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Brandl, N., Seitz, R., Sendtner, N., M\u00fcller, M., and G\u00fclow, K. (2025). Living on the edge: ROS homeostasis in cancer cells and its potential as a therapeutic target. Antioxidants, 14.","DOI":"10.3390\/antiox14081002"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"39948","DOI":"10.1038\/s41598-025-23653-0","article-title":"The dual impact of oxidative stress on breast cancer","volume":"15","author":"Li","year":"2025","journal-title":"Sci. Rep."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1609","DOI":"10.1038\/s41388-025-03336-w","article-title":"ROS\u2013ATM\u2013CHK2 axis stabilizes HIF-1\u03b1 and promotes tumor angiogenesis in hypoxic microenvironment","volume":"44","author":"Bai","year":"2025","journal-title":"Oncogene"},{"key":"ref_37","first-page":"297","article-title":"Oxidative stress in tumor microenvironment\u2014Its role in angiogenesis","volume":"11","author":"Rojas","year":"2008","journal-title":"Chin. J. Lung Cancer"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1186\/s12964-023-01398-5","article-title":"Interplay of oxidative stress, cellular communication and signaling pathways in cancer","volume":"22","author":"Iqbal","year":"2024","journal-title":"Cell Commun. Signal."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Park, M.N., Kim, M., Lee, S., Kang, S., Ahn, C.H., Tallei, T.E., Kim, W., and Kim, B. (2025). Targeting redox signaling through exosomal microRNA: Insights into tumor microenvironment and precision oncology. Antioxidants, 14.","DOI":"10.3390\/antiox14050501"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"621","DOI":"10.1111\/j.1365-2613.2009.00682.x","article-title":"Reactive oxygen species promote angiogenesis in the infarcted rat heart","volume":"90","author":"Zhao","year":"2009","journal-title":"Int. J. Exp. Path."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"625","DOI":"10.1182\/blood-2013-09-512749","article-title":"Oxidative stress in angiogenesis and vascular disease","volume":"123","author":"Kim","year":"2014","journal-title":"Blood"},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Meschiari, G., Minacori, M., Fiorini, S., Tedesco, M., Eufemi, M., and Altieri, F. (2024). Analysis of punicalin and punicalagin interaction with PDIA3 and PDIA1. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms251910531"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"114070","DOI":"10.1016\/j.biopha.2022.114070","article-title":"The synthetic molecule stauprimide impairs cell growth and migration in triple-negative breast cancer","volume":"158","author":"Carrillo","year":"2023","journal-title":"Biomed. Pharmacother."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"429","DOI":"10.1038\/s41586-024-07698-1","article-title":"Tumour vasculature at single-cell resolution","volume":"632","author":"Pan","year":"2024","journal-title":"Nature"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"123121","DOI":"10.1016\/j.lfs.2024.123121","article-title":"Navigating therapeutic prospects by modulating autophagy in colorectal cancer","volume":"358","author":"Rajendran","year":"2024","journal-title":"Life Sci."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/3\/1533\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,2,6]],"date-time":"2026-02-06T05:18:29Z","timestamp":1770355109000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/3\/1533"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,2,4]]},"references-count":45,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2026,2]]}},"alternative-id":["ijms27031533"],"URL":"https:\/\/doi.org\/10.3390\/ijms27031533","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,2,4]]}}}