{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,13]],"date-time":"2026-07-13T18:05:42Z","timestamp":1783965942030,"version":"3.55.0"},"reference-count":45,"publisher":"MDPI AG","issue":"15","license":[{"start":{"date-parts":[[2025,7,25]],"date-time":"2025-07-25T00:00:00Z","timestamp":1753401600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Northwest Cherry Growers, Washington State Fruit Commission","award":["M2201747"],"award-info":[{"award-number":["M2201747"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Dark sweet cherries (DSC) phytochemicals have emerged as a promising dietary strategy to combat triple-negative breast cancer (TNBC). This study explored the effects of DSC extract rich in anthocyanins (ACN) as a chemopreventive agent and as a complement to doxorubicin (DOX) in treating TNBC tumors and metastasis using a 4T1 syngeneic animal model. Initiating ACN intake as a chemopreventive one week before 4T1 cell implantation significantly delayed tumor growth without any signs of toxicity. Both DOX treatment and the combination of DOX-ACN effectively delayed tumor growth rate, but DOX-ACN allowed for body weight gain, which was hindered by DOX alone. As a chemopreventive, ACN downregulated metastasis- and immune-suppression-related genes, including STAT3, Snail1, mTOR, SIRT1, TGF\u03b21, IKK\u03b2, and those unaffected by DOX alone, such as HIF, Cd44, and Rgcc32. Correlations between mRNA levels seen in control and DOX groups were absent in ACN and\/or DOX-ACN groups, indicating that Cd44, mTOR, Rgcc32, SIRT1, Snail1, and TGF\u03b21 may be ACN targets. The DOX-ACN treatment showed a trend toward enhanced efficacy involving CREB, PI3K, Akt-1, and Vim compared to DOX alone. Particularly, ACN significantly suppressed lung metastasis compared to the other groups. ACN also decreased the frequency and incidence of metastasis in the liver, heart, kidneys, and spleen, while their metastatic area (%) and number of breast cancer (BC) metastatic tumor nodules were lowered without reaching significance. Further research is needed to explore the efficacy of combining ACN with drug therapy in the context of drug resistance.<\/jats:p>","DOI":"10.3390\/ijms26157225","type":"journal-article","created":{"date-parts":[[2025,7,25]],"date-time":"2025-07-25T16:06:17Z","timestamp":1753459577000},"page":"7225","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Dark Sweet Cherry Anthocyanins Suppressed Triple-Negative Breast Cancer Pulmonary Metastasis and Downregulated Genes Associated with Metastasis and Therapy Resistance In Vivo"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1605-8744","authenticated-orcid":false,"given":"Ana","family":"Nava-Ochoa","sequence":"first","affiliation":[{"name":"Department of Food Science and Technology, Texas A & M University, College Station, TX 77843, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0746-5949","authenticated-orcid":false,"given":"Lauren W.","family":"Stranahan","sequence":"additional","affiliation":[{"name":"Department of Veterinary Pathobiology, School of Veterinary Medicine & Biomedical Sciences, Texas A & M University, College Station, TX 77843, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8054-6553","authenticated-orcid":false,"given":"Rodrigo","family":"San-Cristobal","sequence":"additional","affiliation":[{"name":"Department of Food Science and Technology, Texas A & M University, College Station, TX 77843, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Susanne U.","family":"Mertens-Talcott","sequence":"additional","affiliation":[{"name":"Department of Food Science and Technology, Texas A & M University, College Station, TX 77843, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1831-7172","authenticated-orcid":false,"given":"Giuliana D.","family":"Noratto","sequence":"additional","affiliation":[{"name":"Department of Food Science and Technology, Texas A & M University, College Station, TX 77843, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,7,25]]},"reference":[{"key":"ref_1","unstructured":"(2025, June 02). World Health Organization, Breast Cancer. Key Facts 2025. Available online: https:\/\/www.who.int\/news-room\/fact-sheets\/detail\/breast-cancer."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Rapoport, B.L., Nayler, S., Mlecnik, B., Smit, T., Heyman, L., Bouquet, I., Martel, M., Galon, J., Benn, C.A., and Anderson, R. (2022). Tumor-Infiltrating Lymphocytes (TILs) in Early Breast Cancer Patients: High CD3, CD8, and Immunoscore Are Associated with a Pathological Complete Response. Cancers, 14.","DOI":"10.3390\/cancers14102525"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1016\/S1470-2045(17)30904-X","article-title":"Tumour-infiltrating lymphocytes and prognosis in different subtypes of breast cancer: A pooled analysis of 3771 patients treated with neoadjuvant therapy","volume":"19","author":"Denkert","year":"2018","journal-title":"Lancet Oncol."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Wu, T.N., Chen, H.M., and Shyur, L.F. (2021). Current Advancements of Plant-Derived Agents for Triple-Negative Breast Cancer Therapy through Deregulating Cancer Cell Functions and Reprogramming Tumor Microenvironment. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms222413571"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1016\/j.jfda.2019.11.001","article-title":"Recent advances in cancer chemoprevention with phytochemicals","volume":"28","author":"Koh","year":"2020","journal-title":"J. Food Drug Anal."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1016\/j.jconrel.2020.12.012","article-title":"A bird\u2019s eye view of the advanced approaches and strategies for overshadowing triple negative breast cancer","volume":"330","author":"Malavia","year":"2021","journal-title":"J. Control. Release"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Lage, N.N., Layosa, M.A.A., Arbizu, S., Chew, B.P., Pedrosa, M.L., Mertens-Talcott, S., Talcott, S., and Noratto, G.D. (2020). Dark sweet cherry (Prunus avium) phenolics enriched in anthocyanins exhibit enhanced activity against the most aggressive breast cancer subtypes without toxicity to normal breast cells. J. Funct. Foods, 64.","DOI":"10.1016\/j.jff.2019.103710"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Silveira Rabelo, A.C., Mertens-Talcott, S.U., Chew, B.P., and Noratto, G. (2022). Dark Sweet Cherry (Prunus avium) Anthocyanins Suppressed ERK1\/2-Akt\/mTOR Cell Signaling and Oxidative Stress: Implications for TNBC Growth and Invasion. Molecules, 27.","DOI":"10.3390\/molecules27217245"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"539","DOI":"10.1007\/s12282-024-01567-5","article-title":"PI3K\/AKT\/mTOR signaling pathway: An important driver and therapeutic target in triple-negative breast cancer","volume":"31","author":"Zhang","year":"2024","journal-title":"Breast Cancer"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"264","DOI":"10.1186\/s12935-019-0986-8","article-title":"Overexpression of CENPF correlates with poor prognosis and tumor bone metastasis in breast cancer","volume":"19","author":"Sun","year":"2019","journal-title":"Cancer Cell Int."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"38","DOI":"10.3389\/fphar.2019.00038","article-title":"Dual Tumor Suppressor and Tumor Promoter Action of Sirtuins in Determining Malignant Phenotype","volume":"10","author":"Carafa","year":"2019","journal-title":"Front. Pharmacol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"101205","DOI":"10.1016\/j.mam.2023.101205","article-title":"Doxorubicin and other anthracyclines in cancers: Activity, chemoresistance and its overcoming","volume":"93","author":"Mattioli","year":"2023","journal-title":"Mol. Aspects Med."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"530","DOI":"10.1186\/s12935-021-02224-z","article-title":"Systematical analysis reveals a strong cancer relevance of CREB1-regulated genes","volume":"21","author":"Zheng","year":"2021","journal-title":"Cancer Cell Int."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Rabelo, A.C.S., Guerreiro, C.A., Shinzato, V.I., Ong, T.P., and Noratto, G. (2023). Anthocyanins Reduce Cell Invasion and Migration through Akt\/mTOR Downregulation and Apoptosis Activation in Triple-Negative Breast Cancer Cells: A Systematic Review and Meta-Analysis. Cancers, 15.","DOI":"10.3390\/cancers15082300"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"To, S.Q., Dmello, R.S., Richards, A.K., Ernst, M., and Chand, A.L. (2022). STAT3 Signaling in Breast Cancer: Multicellular Actions and Therapeutic Potential. Cancers, 14.","DOI":"10.3390\/cancers14020429"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1186\/s12964-020-0527-z","article-title":"Role of STAT3 signaling pathway in breast cancer","volume":"18","author":"Ma","year":"2020","journal-title":"Cell Commun. Signal"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1016\/j.semcancer.2022.08.003","article-title":"STAT family of transcription factors in breast cancer: Pathogenesis and therapeutic opportunities and challenges","volume":"86","author":"Wong","year":"2022","journal-title":"Semin. Cancer Biol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"154676","DOI":"10.1016\/j.prp.2023.154676","article-title":"Hypoxia inducible factor-1alpha (HIF-1alpha) in breast cancer: The crosstalk with oncogenic and onco-suppressor factors in regulation of cancer hallmarks","volume":"248","author":"Mirzaei","year":"2023","journal-title":"Pathol. Res. Pract."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"174","DOI":"10.1007\/s12032-023-02037-2","article-title":"Oncogenic metabolic reprogramming in breast cancer: Focus on signaling pathways and mitochondrial genes","volume":"40","author":"Malayil","year":"2023","journal-title":"Med. Oncol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.semcancer.2019.12.008","article-title":"Natural products targeting the PI3K-Akt-mTOR signaling pathway in cancer: A novel therapeutic strategy","volume":"80","author":"Tewari","year":"2022","journal-title":"Semin. Cancer Biol."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Onyiba, C.I., Scarlett, C.J., and Weidenhofer, J. (2022). The Mechanistic Roles of Sirtuins in Breast and Prostate Cancer. Cancers, 14.","DOI":"10.3390\/cancers14205118"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"104880","DOI":"10.1016\/j.phrs.2020.104880","article-title":"Emerging role of sirtuins in breast cancer metastasis and multidrug resistance: Implication for novel therapeutic strategies targeting sirtuins","volume":"158","author":"Sinha","year":"2020","journal-title":"Pharmacol. Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"121011","DOI":"10.1016\/j.lfs.2022.121011","article-title":"TGF-beta at the crossroads of multiple prognosis in breast cancer, and beyond","volume":"310","author":"Shukla","year":"2022","journal-title":"Life Sci."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"504","DOI":"10.1159\/000531802","article-title":"Targeting Breast Cancer Signaling via Phytomedicine and Nanomedicine","volume":"108","author":"Ansari","year":"2023","journal-title":"Pharmacology"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1023\/B:CLIN.0000037705.25256.d3","article-title":"Transforming growth factor-beta facilitates breast carcinoma metastasis by promoting tumor cell survival","volume":"21","author":"Yu","year":"2004","journal-title":"Clin. Exp. Metastasis"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1782","DOI":"10.7150\/ijbs.23586","article-title":"Novel CD44-downstream signaling pathways mediating breast tumor invasion","volume":"14","author":"Ouhtit","year":"2018","journal-title":"Int. J. Biol. Sci."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"234","DOI":"10.1158\/1055-9965.EPI-13-0785","article-title":"Molecular biomarkers of cancer stem\/progenitor cells associated with progression, metastases, and treatment resistance of aggressive cancers","volume":"23","author":"Mimeault","year":"2014","journal-title":"Cancer Epidemiol. Biomarkers Prev."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1","DOI":"10.3233\/BD-190409","article-title":"Role of CD44 in breast cancer","volume":"39","author":"Alhendi","year":"2020","journal-title":"Breast Dis."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"342","DOI":"10.1186\/s13046-023-02928-2","article-title":"RGCC-mediated PLK1 activity drives breast cancer lung metastasis by phosphorylating AMPKalpha2 to activate oxidative phosphorylation and fatty acid oxidation","volume":"42","author":"Cheng","year":"2023","journal-title":"J. Exp. Clin. Cancer Res."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"617","DOI":"10.1161\/CIRCULATIONAHA.108.841502","article-title":"Response gene to complement 32, a novel hypoxia-regulated angiogenic inhibitor","volume":"120","author":"An","year":"2009","journal-title":"Circulation"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1016\/j.advenzreg.2006.01.004","article-title":"Roles of the RAF\/MEK\/ERK and PI3K\/PTEN\/AKT pathways in malignant transformation and drug resistance","volume":"46","author":"McCubrey","year":"2006","journal-title":"Adv. Enzyme Regul."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"111068","DOI":"10.1016\/j.biopha.2020.111068","article-title":"Regulatory mechanisms and clinical significance of vimentin in breast cancer","volume":"133","author":"Chen","year":"2021","journal-title":"Biomed. Pharmacother."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"418","DOI":"10.1016\/j.pharmthera.2013.02.004","article-title":"Polarity proteins as regulators of cell junction complexes: Implications for breast cancer","volume":"138","author":"Bazzoun","year":"2013","journal-title":"Pharmacol. Ther."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"113128","DOI":"10.1016\/j.biopha.2022.113128","article-title":"The anticancer effects of cyanidin 3-O-glucoside combined with 5-fluorouracil on lung large-cell carcinoma in nude mice","volume":"151","author":"Wu","year":"2022","journal-title":"Biomed. Pharmacother."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"153227","DOI":"10.1016\/j.phymed.2020.153227","article-title":"Breast Tumor Microenvironment: Emerging target of therapeutic phytochemicals","volume":"70","author":"Malla","year":"2020","journal-title":"Phytomedicine"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1595","DOI":"10.1007\/s00210-023-02429-1","article-title":"Preventive and therapeutic use of herbal compounds against doxorubicin induced hepatotoxicity: A comprehensive review","volume":"396","author":"Mahmoudi","year":"2023","journal-title":"Naunyn-Schmiedeb. Arch. Pharmacol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1141","DOI":"10.6026\/973206300181141","article-title":"Oxidative stress in breast cancer after chemotherapy","volume":"18","author":"Mannangatti","year":"2022","journal-title":"Bioinformation"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"374","DOI":"10.1097\/COC.0000000000001013","article-title":"Cardiac Metastatic Tumors: Current Knowledge","volume":"46","author":"Kopcik","year":"2023","journal-title":"Am. J. Clin. Oncol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"750186","DOI":"10.3389\/fcvm.2021.750186","article-title":"Luteolin Prevents Cardiac Dysfunction and Improves the Chemotherapeutic Efficacy of Doxorubicin in Breast Cancer","volume":"8","author":"Shi","year":"2021","journal-title":"Front. Cardiovasc. Med."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"15593258221120485","DOI":"10.1177\/15593258221120485","article-title":"Phytochemicals That Interfere With Drug Metabolism and Transport, Modifying Plasma Concentration in Humans and Animals","volume":"20","year":"2022","journal-title":"Dose Response"},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Nava-Ochoa, A., Mertens-Talcott, S.U., Talcott, S.T., and Noratto, G.D. (2025). Dark Sweet Cherry (L.) Juice Phenolics Rich in Anthocyanins Exhibit Potential to Inhibit Drug Resistance Mechanisms in 4T1 Breast Cancer Cells via the Drug Metabolism Pathway. Curr. Issues Mol. Biol., 47.","DOI":"10.3390\/cimb47030213"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1186\/s13058-015-0558-3","article-title":"Granulocytic immune infiltrates are essential for the efficient formation of breast cancer liver metastases","volume":"17","author":"Tabaries","year":"2015","journal-title":"Breast Cancer Res."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Giusti, M.M., and Wrolstad, R.E. (2001). Characterization and Measurement of Anthocyanins by UV-Visible Spectroscopy. Curr. Protoc. Food Anal. Chem., F1.2.1\u2013F1.2.13.","DOI":"10.1002\/0471142913.faf0102s00"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1101","DOI":"10.1038\/nprot.2008.73","article-title":"Analyzing real-time PCR data by the comparative C(T) method","volume":"3","author":"Schmittgen","year":"2008","journal-title":"Nat. Protoc."},{"key":"ref_45","first-page":"54040","article-title":"An Orthotopic Mouse Model of Spontaneous Breast Cancer Metastasis","volume":"114","author":"Paschall","year":"2016","journal-title":"J. Vis. Exp."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/15\/7225\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:16:21Z","timestamp":1760033781000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/15\/7225"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,7,25]]},"references-count":45,"journal-issue":{"issue":"15","published-online":{"date-parts":[[2025,8]]}},"alternative-id":["ijms26157225"],"URL":"https:\/\/doi.org\/10.3390\/ijms26157225","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,7,25]]}}}