{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,11]],"date-time":"2026-03-11T12:41:42Z","timestamp":1773232902595,"version":"3.50.1"},"reference-count":57,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2021,2,21]],"date-time":"2021-02-21T00:00:00Z","timestamp":1613865600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Catalysis S.L","award":["2016.0036"],"award-info":[{"award-number":["2016.0036"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Nutrients"],"abstract":"<jats:p>Whereas the prevalence of several cancer types is decreasing, skin malignancies are growing more common every year. Malignant melanoma is the most aggressive form of skin cancer with high metastatic capacity. In most cases, malignant melanoma shows acquired therapy resistance. We evaluated the ability of Ocoxin, a natural compound-based antioxidant and anti-inflammatory nutritional complement, to exert an antitumor effect in melanoma. To do so, the cytotoxicity of Ocoxin in a panel of BRAF-mutated murine and human melanoma cell lines was tested alone and in combination with BRAF inhibitor Vemurafenib. Our results revealed a potent cytotoxic effect of Ocoxin against melanoma cells and a synergic effect when combined with Vemurafenib, reducing viability and increasing apoptosis. Besides, Ocoxin interferes with the cell cycle, impairs the inherent and fibroblast-mediated melanoma cell migration, and reduces resistance to BRAF inhibition. Proteomic analysis revealed reduced tumor secretion of inflammatory factors Galectin-1, Osteopontin, CCL5, and CCL9 upon treatment with Ocoxin. Moreover, RNASeq showed that Ocoxin downregulated the cell cycle and proliferation-related genes. In vivo, Ocoxin reduced the number of lung metastasis of YUMM-1.7 melanoma cells. Therefore, Ocoxin arises as a good candidate for clinical trials analyzing the beneficial effects in patients suffering from this cutaneous malignancy.<\/jats:p>","DOI":"10.3390\/nu13020686","type":"journal-article","created":{"date-parts":[[2021,2,21]],"date-time":"2021-02-21T22:04:15Z","timestamp":1613945055000},"page":"686","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["Ocoxin Increases the Antitumor Effect of BRAF Inhibition and Reduces Cancer Associated Fibroblast-Mediated Chemoresistance and Protumoral Activity in Metastatic Melanoma"],"prefix":"10.3390","volume":"13","author":[{"given":"Aitor","family":"Benedicto","sequence":"first","affiliation":[{"name":"Department of Cell Biology and Histology, School of Medicine and Nursing, University of the Basque Country, 48940 Leioa, Bizkaia, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Iera","family":"Hernandez-Unzueta","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Histology, School of Medicine and Nursing, University of the Basque Country, 48940 Leioa, Bizkaia, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eduardo","family":"Sanz","sequence":"additional","affiliation":[{"name":"Research and Development, Catalysis S.L., 28016 Madrid, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6429-615X","authenticated-orcid":false,"given":"Joana","family":"M\u00e1rquez","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Histology, School of Medicine and Nursing, University of the Basque Country, 48940 Leioa, Bizkaia, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,2,21]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"949","DOI":"10.1038\/nature00766","article-title":"Mutations of the BRAF Gene in Human Cancer","volume":"417","author":"Davies","year":"2002","journal-title":"Nature"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1016\/j.cell.2018.03.035","article-title":"Oncogenic Signaling Pathways in the Cancer Genome Atlas","volume":"173","author":"Mina","year":"2018","journal-title":"Cell"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1097\/CCO.0b013e3282f5271c","article-title":"The MAPK Pathway in Melanoma","volume":"20","author":"Fecher","year":"2008","journal-title":"Curr. Opin. Oncol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"313","DOI":"10.1016\/j.bbrc.2010.07.092","article-title":"RAF Protein-Serine\/Threonine Kinases: Structure and Regulation","volume":"399","author":"Roskoski","year":"2010","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Hamid, O., Cowey, C.L., Offner, M., Faries, M., and Carvajal, R.D. (2019). Efficacy, Safety, and Tolerability of Approved Combination BRAF and MEK Inhibitor Regimens for BRAF-Mutant Melanoma. Cancers, 11.","DOI":"10.3390\/cancers11111642"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/j.phymed.2017.12.001","article-title":"Genistein Induced Anticancer Effects on Pancreatic Cancer Cell Lines Involves Mitochondrial Apoptosis, G(0)\/G(1)Cell Cycle Arrest and Regulation of STAT3 Signalling Pathway","volume":"39","author":"Bi","year":"2018","journal-title":"Phytomedicine"},{"key":"ref_7","first-page":"634","article-title":"Curcumin Promoted miR-34a Expression and Suppressed Proliferation of Gastric Cancer Cells","volume":"34","author":"Sun","year":"2019","journal-title":"Cancer Biother. Radiopharm."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"925640","DOI":"10.1155\/2015\/925640","article-title":"Implications of Green Tea and its Constituents in the Prevention of Cancer Via the Modulation of Cell Signalling Pathway","volume":"2015","author":"Rahmani","year":"2015","journal-title":"Biomed. Res. Int."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"555","DOI":"10.1097\/MPA.0000000000001277","article-title":"Ocoxin Oral Solution Exerts an Antitumoral Effect in Pancreatic Cancer and Reduces the Stromal-Mediated Chemoresistance","volume":"48","author":"Benedicto","year":"2019","journal-title":"Pancreas"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Marquez, J., Mena, J., Hernandez-Unzueta, I., Benedicto, A., Sanz, E., Arteta, B., and Olaso, E. (2015). Ocoxin(R) Oral Solution Slows Down Tumor Growth in an Experimental Model of Colorectal Cancer Metastasis to the Liver in Balb\/C Mice. Oncol. Rep.","DOI":"10.3892\/or.2015.4486"},{"key":"ref_11","first-page":"9719730","article-title":"Ocoxin Modulates Cancer Stem Cells and M2 Macrophage Polarization in Glioblastoma","volume":"2019","author":"Gargini","year":"2019","journal-title":"Oxidative Med. Cell. Longev."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"6231","DOI":"10.18632\/oncotarget.6862","article-title":"Antitumoral Effect of Ocoxin on Acute Myeloid Leukemia","volume":"7","author":"Sanz","year":"2016","journal-title":"Oncotarget"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1159","DOI":"10.1080\/01635581.2015.1068819","article-title":"Effect of Oncoxin Oral Solution in HER2-Overexpressing Breast Cancer","volume":"67","author":"Gonzalez","year":"2015","journal-title":"Nutr. Cancer"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"172","DOI":"10.3329\/taj.v22i2.37718","article-title":"Findings of the 3-Month Supportive Treatment with Oncoxin Solution Beside the Standard Modalities of Patients with Different Neoplastic Diseases","volume":"22","author":"Islam","year":"2009","journal-title":"TAJ J. Teach. Assoc."},{"key":"ref_15","first-page":"463","article-title":"Oncoxin Nutritional Supplement in the Management of Chemotherapy- and\/Or Radiotherapy-Associated Oral Mucositis","volume":"10","author":"Shumsky","year":"2019","journal-title":"Mol. Clin. Oncol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"317","DOI":"10.4172\/1948-5956.1000562","article-title":"Oncoxin-Viusid with Radiotherapy and Chemotherapy in Patients withHead and Neck Cancer: Results from a Phase II, Randomised, Double-Blind Study","volume":"10","author":"Rivas","year":"2018","journal-title":"J. Cancer Sci. Ther."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"4002","DOI":"10.3892\/ol.2017.6016","article-title":"Ocoxin Oral Solution((R)) as a Complement to Irinotecan Chemotherapy in the Metastatic Progression of Colorectal Cancer to the Liver","volume":"13","author":"Benedicto","year":"2017","journal-title":"Oncol. Lett."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"281","DOI":"10.3389\/fonc.2020.00281","article-title":"Metabolic Plasticity in Chemotherapy Resistance","volume":"10","author":"Desbats","year":"2020","journal-title":"Front. Oncol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1007\/978-3-030-34025-4_2","article-title":"Tumor Microenvironment\u2014Selective Pressures Boosting Cancer Progression","volume":"1219","author":"Nunes","year":"2020","journal-title":"Adv. Exp. Med. Biol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1016\/j.canlet.2016.01.043","article-title":"Tumor Microenvironment and Therapeutic Response","volume":"387","author":"Wu","year":"2017","journal-title":"Cancer Lett."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2412","DOI":"10.1089\/ten.tea.2013.0473","article-title":"Fibroblasts Protect Melanoma Cells from the Cytotoxic Effects of Doxorubicin","volume":"20","author":"Tiago","year":"2014","journal-title":"Tissue Eng. Part A"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"14235","DOI":"10.1007\/s13277-016-5293-6","article-title":"Targeted Deactivation of Cancer-Associated Fibroblasts by \u0392-Catenin Ablation Suppresses Melanoma Growth","volume":"37","author":"Zhou","year":"2016","journal-title":"Tumour Biol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"29","DOI":"10.3727\/096504015X14452563486011","article-title":"Curcumin Enhances the Effects of 5-Fluorouracil and Oxaliplatin in Inducing Gastric Cancer Cell Apoptosis both in Vitro and in Vivo","volume":"23","author":"Zhou","year":"2016","journal-title":"Oncol. Res."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"674","DOI":"10.1053\/jhep.2003.50068","article-title":"Proangiogenic Role of Tumor-Activated Hepatic Stellate Cells in Experimental Melanoma Metastasis","volume":"37","author":"Olaso","year":"2003","journal-title":"Hepatology"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"250","DOI":"10.1038\/nature17392","article-title":"sFRP2 in the Aged Microenvironment Drives Melanoma Metastasis and Therapy Resistance","volume":"532","author":"Kaur","year":"2016","journal-title":"Nature"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"841","DOI":"10.1016\/j.cell.2018.01.009","article-title":"CD10(+)GPR77(+) Cancer-Associated Fibroblasts Promote Cancer Formation and Chemoresistance by Sustaining Cancer Stemness","volume":"172","author":"Su","year":"2018","journal-title":"Cell"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"3932","DOI":"10.7150\/thno.25541","article-title":"Carcinoma-Associated Fibroblasts Promote the Stemness and Chemoresistance of Colorectal Cancer by Transferring Exosomal lncRNA H19","volume":"8","author":"Ren","year":"2018","journal-title":"Theranostics"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"500","DOI":"10.1038\/nature11183","article-title":"Tumour Micro-Environment Elicits Innate Resistance to RAF Inhibitors through HGF Secretion","volume":"487","author":"Straussman","year":"2012","journal-title":"Nature"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1097\/PPO.0000000000000237","article-title":"Melanoma Brain Metastases: Current Areas of Investigation and Future Directions","volume":"23","author":"Tawbi","year":"2017","journal-title":"Cancer J."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Kee, J.Y., Han, Y.H., Kim, D.S., Mun, J.G., Park, S.H., So, H.S., Park, S.J., Park, R., Um, J.Y., and Hong, S.H. (2017). \u0392-Lapachone Suppresses the Lung Metastasis of Melanoma Via the MAPK Signaling Pathway. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0176937"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"323","DOI":"10.1016\/S1470-2045(14)70012-9","article-title":"Safety and Efficacy of Vemurafenib in BRAF(V600E) and BRAF(V600K) Mutation-Positive Melanoma (BRIM-3): Extended Follow-Up of a Phase 3, Randomised, Open-Label Study","volume":"15","author":"McArthur","year":"2014","journal-title":"Lancet Oncol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1016\/j.ejca.2015.09.013","article-title":"Survival of Patients with Advanced Metastatic Melanoma: The Impact of Novel Therapies","volume":"53","author":"Ugurel","year":"2016","journal-title":"Eur. J. Cancer"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2507","DOI":"10.1056\/NEJMoa1103782","article-title":"Improved Survival with Vemurafenib in Melanoma with BRAF V600E Mutation","volume":"364","author":"Chapman","year":"2011","journal-title":"N. Engl. J. Med."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"358","DOI":"10.1016\/S0140-6736(12)60868-X","article-title":"Dabrafenib in BRAF-Mutated Metastatic Melanoma: A Multicentre, Open-Label, Phase 3 Randomised Controlled Trial","volume":"380","author":"Hauschild","year":"2012","journal-title":"Lancet"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1104","DOI":"10.15252\/emmm.201404914","article-title":"Intra- and Inter-Tumor Heterogeneity in a Vemurafenib-Resistant Melanoma Patient and Derived Xenografts","volume":"7","author":"Kemper","year":"2015","journal-title":"EMBO Mol. Med."},{"key":"ref_36","first-page":"113","article-title":"Antitumoral Effect of Ocoxin, a Natural Compound-Containing Nutritional Supplement, in Small Cell Lung Cancer","volume":"53","author":"Sanz","year":"2018","journal-title":"Int. J. Oncol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"450S","DOI":"10.1080\/07315724.2001.10719184","article-title":"Scientific Rationale for using High-Dose Multiple Micronutrients as an Adjunct to Standard and Experimental Cancer Therapies","volume":"20","author":"Prasad","year":"2001","journal-title":"J. Am. Coll. Nutr."},{"key":"ref_38","first-page":"240","article-title":"miR-490-5p Regulates the Proliferation, Migration, Invasion and Epithelial-mesenchymal Transition of Pharyngolaryngeal Cancer Cells by Targeting Mitogen-activated Protein Kinase Kinasekinase 9","volume":"44","author":"Abdeyrim","year":"2019","journal-title":"Int. J. Mol. Med."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Chuang, H.C., and Tan, T.H. (2019). MAP4K Family Kinases and DUSP Family Phosphatases in T-Cell Signaling and Systemic Lupus Erythematosus. Cells, 8.","DOI":"10.3390\/cells8111433"},{"key":"ref_40","first-page":"6497","article-title":"MicroRNA-148a Regulates the MAPK\/ERK Signaling Pathway and Suppresses the Development of Esophagus Squamous Cell Carcinoma Via Targeting MAP3K9","volume":"23","author":"Zhang","year":"2019","journal-title":"Eur. Rev. Med. Pharmacol. Sci."},{"key":"ref_41","first-page":"4288","article-title":"MicroRNA-98-5p Regulates the Proliferation and Apoptosis of A549 Cells by Targeting MAP4K3","volume":"18","author":"Wang","year":"2019","journal-title":"Oncol. Lett."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1016\/j.omtn.2018.08.012","article-title":"miR-7 Suppresses Tumor Progression by Directly Targeting MAP3K9 in Pancreatic Cancer","volume":"13","author":"Xia","year":"2018","journal-title":"Mol. Ther. Nucleic Acids"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"876","DOI":"10.1080\/02713680802382989","article-title":"Inhibitor of Apoptosis Proteins Gene Expression and its Correlation with Prognostic Factors in Primary and Metastatic Uveal Melanoma","volume":"33","author":"Lederman","year":"2008","journal-title":"Curr. Eye Res."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s41598-020-72864-0","article-title":"Transcriptional Characterization of Conjunctival Melanoma Identifies the Cellular Tumor Microenvironment and Prognostic Gene Signatures","volume":"10","author":"Wolf","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"369","DOI":"10.29252\/ibj.23.6.369","article-title":"BIRC5 Gene Disruption Via CRISPR\/Cas9n Platform Suppress Acute Myelocytic Leukemia Progression","volume":"23","author":"Narimani","year":"2019","journal-title":"Iran. Biomed. J."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1965","DOI":"10.1002\/mc.22654","article-title":"Vitamin C at High Concentrations Induces Cytotoxicity in Malignant Melanoma but Promotes Tumor Growth at Low Concentrations","volume":"56","author":"Yang","year":"2017","journal-title":"Mol. Carcinog."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1163","DOI":"10.1158\/1535-7163.MCT-15-0973","article-title":"A Novel Plant Sesquiterpene Lactone Derivative, DETD-35, Suppresses BRAFV600E Mutant Melanoma Growth and Overcomes Acquired Vemurafenib Resistance in Mice","volume":"15","author":"Feng","year":"2016","journal-title":"Mol. Cancer Ther."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s13045-016-0237-6","article-title":"Autologous Cytokine-Induced Killer Cell Transfusion Increases overall Survival in Advanced Pancreatic Cancer","volume":"9","author":"Wang","year":"2016","journal-title":"J. Hematol. Oncol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1186\/s13045-015-0185-6","article-title":"Advances in the Management of Gastrointestinal Cancers--an Upcoming Role of Immune Checkpoint Blockade","volume":"8","author":"Goel","year":"2015","journal-title":"J. Hematol. Oncol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s13045-015-0141-5","article-title":"Novel Approaches in the Management of Pancreatic Ductal Adenocarcinoma: Potential Promises for the Future","volume":"8","author":"Goel","year":"2015","journal-title":"J. Hematol. Oncol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"11924","DOI":"10.1038\/srep11924","article-title":"TGF\u03b21 Secreted by Cancer-Associated Fibroblasts Induces Epithelial-Mesenchymal Transition of Bladder Cancer Cells through lncRNA-ZEB2NAT","volume":"5","author":"Zhuang","year":"2015","journal-title":"Sci. Rep."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"1041","DOI":"10.1093\/carcin\/bgw093","article-title":"TGF-Beta-Induced Stromal CYR61 Promotes Resistance to Gemcitabine in Pancreatic Ductal Adenocarcinoma through Downregulation of the Nucleoside Transporters hENT1 and hCNT3","volume":"37","author":"Hesler","year":"2016","journal-title":"Carcinogenesis"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"4821","DOI":"10.1038\/onc.2014.410","article-title":"Osteopontin Mediates an MZF1-TGF-\u03921-Dependent Transformation of Mesenchymal Stem Cells into Cancer-Associated Fibroblasts in Breast Cancer","volume":"34","author":"Weber","year":"2015","journal-title":"Oncogene"},{"key":"ref_54","doi-asserted-by":"crossref","unstructured":"Aldinucci, D., Borghese, C., and Casagrande, N. (2020). The CCL5\/CCR5 Axis in Cancer Progression. Cancers, 12.","DOI":"10.3390\/cancers12071765"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"2000","DOI":"10.1016\/j.celrep.2016.04.084","article-title":"Reprogramming Tumor-Associated Macrophages by Antibody Targeting Inhibits Cancer Progression and Metastasis","volume":"15","author":"Georgoudaki","year":"2016","journal-title":"Cell Rep."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"497","DOI":"10.1038\/nm.4082","article-title":"Genotype Tunes Pancreatic Ductal Adenocarcinoma Tissue Tension to Induce Matricellular Fibrosis and Tumor Progression","volume":"22","author":"Laklai","year":"2016","journal-title":"Nat. Med."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"462","DOI":"10.1038\/nm.4099","article-title":"Regulation of Pancreatic Cancer Aggressiveness by Stromal Stiffening","volume":"22","author":"Rath","year":"2016","journal-title":"Nat. Med."}],"container-title":["Nutrients"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6643\/13\/2\/686\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T05:27:19Z","timestamp":1760160439000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6643\/13\/2\/686"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,2,21]]},"references-count":57,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2021,2]]}},"alternative-id":["nu13020686"],"URL":"https:\/\/doi.org\/10.3390\/nu13020686","relation":{},"ISSN":["2072-6643"],"issn-type":[{"value":"2072-6643","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,2,21]]}}}