{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,10]],"date-time":"2026-07-10T19:49:36Z","timestamp":1783712976730,"version":"3.55.0"},"reference-count":42,"publisher":"MDPI AG","issue":"24","license":[{"start":{"date-parts":[[2022,12,17]],"date-time":"2022-12-17T00:00:00Z","timestamp":1671235200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Autophagy is a lysosomal degradation and recycling process involved in tumor progression and drug resistance. The aim of this work was to inhibit autophagy and increase apoptosis in a 3D model of human colorectal cancer by combined treatment with our patented natural product Prunus spinosa + nutraceutical activator complex (PsT + NAC\u00ae) and 5-fluorouracil (5-FU). By means of cytotoxic evaluation (MTT assay), cytofluorimetric analysis, light and fluorescence microscopy investigation and Western blotting evaluation of the molecular pathway PI3\/AKT\/mTOR, Caspase-9, Caspase-3, Beclin1, p62 and LC3, we demonstrated that the combination PsT + NAC\u00ae and 5-FU significantly reduces autophagy by increasing the apoptotic phenomenon. These results demonstrate the importance of using non-toxic natural compounds to improve the therapeutic efficacy and reduce the side effects induced by conventional drugs in human colon cancer.<\/jats:p>","DOI":"10.3390\/ijms232416098","type":"journal-article","created":{"date-parts":[[2022,12,19]],"date-time":"2022-12-19T05:25:46Z","timestamp":1671427546000},"page":"16098","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Prunus spinosa Extract Sensitized HCT116 Spheroids to 5-Fluorouracil Toxicity, Inhibiting Autophagy"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2802-9142","authenticated-orcid":false,"given":"Maria","family":"Condello","sequence":"first","affiliation":[{"name":"National Center for Drug Research and Evaluation, National Institute of Health, 00161 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4501-8661","authenticated-orcid":false,"given":"Rosa","family":"Vona","sequence":"additional","affiliation":[{"name":"Center for Gender-Specific Medicine, National Institute of Health, 00161 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9079-1181","authenticated-orcid":false,"given":"Stefania","family":"Meschini","sequence":"additional","affiliation":[{"name":"National Center for Drug Research and Evaluation, National Institute of Health, 00161 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,12,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"209","DOI":"10.3322\/caac.21660","article-title":"Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries","volume":"71","author":"Sung","year":"2021","journal-title":"CA A Cancer J. Clin."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"111285","DOI":"10.1016\/j.biopha.2021.111285","article-title":"5-Fluorouracil (5-FU) Resistance and the New Strategy to Enhance the Sensitivity against Cancer: Implication of DNA Repair Inhibition","volume":"137","author":"Sethy","year":"2021","journal-title":"Biomed. Pharmacother."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.clcc.2014.11.002","article-title":"A Review of the Evolution of Systemic Chemotherapy in the Management of Colorectal Cancer","volume":"14","author":"Gustavsson","year":"2015","journal-title":"Clin. Color. Cancer"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"107447","DOI":"10.1016\/j.pharmthera.2019.107447","article-title":"5-Fluorouracil and Other Fluoropyrimidines in Colorectal Cancer: Past, Present and Future","volume":"206","author":"Vodenkova","year":"2020","journal-title":"Pharmacol. Ther."},{"key":"ref_5","first-page":"141","article-title":"Drug Resistance and Combating Drug Resistance in Cancer","volume":"2","author":"Wang","year":"2019","journal-title":"CDR"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1038\/s41586-019-1730-1","article-title":"A View on Drug Resistance in Cancer","volume":"575","author":"Vasan","year":"2019","journal-title":"Nature"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Fontana, F., Anselmi, M., and Limonta, P. (2022). Molecular Mechanisms of Cancer Drug Resistance: Emerging Biomarkers and Promising Targets to Overcome Tumor Progression. Cancers, 14.","DOI":"10.3390\/cancers14071614"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"891652","DOI":"10.3389\/fonc.2022.891652","article-title":"Multidrug Resistance in Cancer: Understanding Molecular Mechanisms, Immunoprevention and Therapeutic Approaches","volume":"12","author":"Emran","year":"2022","journal-title":"Front. Oncol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"7032614","DOI":"10.1155\/2022\/7032614","article-title":"Research Progress and Prospects of Autophagy in the Mechanism of Multidrug Resistance in Tumors","volume":"2022","author":"Long","year":"2022","journal-title":"J. Oncol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1186\/s13045-020-01000-2","article-title":"Targeting Autophagy to Overcome Drug Resistance: Further Developments","volume":"13","author":"Chang","year":"2020","journal-title":"J. Hematol. Oncol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"538","DOI":"10.1007\/s10637-020-01032-y","article-title":"Autophagy Modulating Agents as Chemosensitizers for Cisplatin Therapy in Cancer","volume":"39","author":"Piska","year":"2021","journal-title":"Investig. New Drugs"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1186\/s12943-020-1138-4","article-title":"Autophagy and Autophagy-Related Proteins in Cancer","volume":"19","author":"Li","year":"2020","journal-title":"Mol. Cancer"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Azwar, S., Seow, H.F., Abdullah, M., Faisal Jabar, M., and Mohtarrudin, N. (2021). Recent Updates on Mechanisms of Resistance to 5-Fluorouracil and Reversal Strategies in Colon Cancer Treatment. Biology, 10.","DOI":"10.3390\/biology10090854"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"658636","DOI":"10.3389\/fonc.2021.658636","article-title":"5-Fluorouracil: A Narrative Review on the Role of Regulatory Mechanisms in Driving Resistance to This Chemotherapeutic Agent","volume":"11","author":"Abak","year":"2021","journal-title":"Front. Oncol."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Sasaki, K., Tsuno, N.H., Sunami, E., Tsurita, G., Kawai, K., Okaji, Y., Nishikawa, T., Shuno, Y., Hongo, K., and Hiyoshi, M. (2010). Chloroquine Potentiates the Anti-Cancer Effect of 5-Fluorouracil on Colon Cancer Cells. BMC Cancer, 10.","DOI":"10.1186\/1471-2407-10-370"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"675","DOI":"10.1097\/CAD.0b013e328353f8c7","article-title":"Resistance of Colon Cancer to 5-Fluorouracil May Be Overcome by Combination with Chloroquine, an in Vivo Study","volume":"23","author":"Sasaki","year":"2012","journal-title":"Anti-Cancer Drugs"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"190","DOI":"10.1186\/s13046-017-0661-7","article-title":"Curcumin Synergizes with 5-Fluorouracil by Impairing AMPK\/ULK1-Dependent Autophagy, AKT Activity and Enhancing Apoptosis in Colon Cancer Cells with Tumor Growth Inhibition in Xenograft Mice","volume":"36","author":"Zhang","year":"2017","journal-title":"J. Exp. Clin. Cancer Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1016\/j.sajb.2019.02.006","article-title":"Comparative Assessment of Prunus Spinosa L. Flower Extract in Non-Neoplastic Hepatocytes and Hepatoblastoma Cells","volume":"123","author":"Murati","year":"2019","journal-title":"S. Afr. J. Bot."},{"key":"ref_19","first-page":"297","article-title":"Antioxidant Activity and Cytotoxic Effects of Prunus Spinosa L. Fruit Extract on Various Cancer Cell Lines","volume":"34","author":"Karakas","year":"2019","journal-title":"MMJ"},{"key":"ref_20","unstructured":"Meschini, S., and Mastrodonato, F. (2015). Estratti di Prunus Spinosa ad Attivit\u00e0 Antitumorale. (No. RM2015A 000133), Italian Patent."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Meschini, S., Pellegrini, E., Condello, M., Occhionero, G., Delfine, S., Condello, G., and Mastrodonato, F. (2017). Cytotoxic and Apoptotic Activities of Prunus Spinosa Trigno Ecotype Extract on Human Cancer Cells. Molecules, 22.","DOI":"10.3390\/molecules22091578"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"109281","DOI":"10.1016\/j.biopha.2019.109281","article-title":"Anticancer Activity of \u201cTrigno M\u201d, Extract of Prunus Spinosa Drupes, against in Vitro 3D and in Vivo Colon Cancer Models","volume":"118","author":"Condello","year":"2019","journal-title":"Biomed. Pharmacother."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"3142","DOI":"10.1111\/cas.14532","article-title":"5-Fluorouracil Resistance Mechanisms in Colorectal Cancer: From Classical Pathways to Promising Processes","volume":"111","author":"Blondy","year":"2020","journal-title":"Cancer Sci."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Yun, C.W., and Lee, S.H. (2018). The Roles of Autophagy in Cancer. Int. J. Mol. Sci., 19.","DOI":"10.3390\/ijms19113466"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"142","DOI":"10.1186\/s12935-022-02561-7","article-title":"A Systematic Review of the Therapeutic Effects of Resveratrol in Combination with 5-Fluorouracil during Colorectal Cancer Treatment: With a Special Focus on the Oxidant, Apoptotic, and Anti-Inflammatory Activities","volume":"22","author":"Moutabian","year":"2022","journal-title":"Cancer Cell Int."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"126","DOI":"10.1186\/s12935-020-01206-x","article-title":"Herbal Melanin Inhibits Colorectal Cancer Cell Proliferation by Altering Redox Balance, Inducing Apoptosis, and Modulating MAPK Signaling","volume":"20","author":"AlHaidan","year":"2020","journal-title":"Cancer Cell Int."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Condello, M., and Meschini, S. (2021). Role of Natural Antioxidant Products in Colorectal Cancer Disease: A Focus on a Natural Compound Derived from Prunus Spinosa, Trigno Ecotype. Cells, 10.","DOI":"10.3390\/cells10123326"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"101051","DOI":"10.1016\/j.tranon.2021.101051","article-title":"The Remodelling of Actin Composition as a Hallmark of Cancer","volume":"14","author":"Suresh","year":"2021","journal-title":"Transl. Oncol"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Uray, I.P., and Uray, K. (2021). Mechanotransduction at the Plasma Membrane-Cytoskeleton Interface. IJMS, 22.","DOI":"10.3390\/ijms222111566"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"eaau9433","DOI":"10.1126\/sciadv.aau9433","article-title":"Efficient Apoptosis Requires Feedback Amplification of Upstream Apoptotic Signals by Effector Caspase-3 or -7","volume":"5","author":"McComb","year":"2019","journal-title":"Sci. Adv."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"76","DOI":"10.1016\/j.neo.2021.12.003","article-title":"MLH1 Mediates Cytoprotective Nucleophagy to Resist 5-Fluorouracil-Induced Cell Death in Colorectal Carcinoma","volume":"24","author":"Manzoor","year":"2022","journal-title":"Neoplasia"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1684","DOI":"10.1080\/10717544.2022.2079771","article-title":"Recent Advances in Anti-Multidrug Resistance for Nano-Drug Delivery System","volume":"29","author":"Wang","year":"2022","journal-title":"Drug Deliv."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"13259","DOI":"10.2147\/CMAR.S279672","article-title":"Regulation of Autophagy by Glycolysis in Cancer","volume":"12","author":"Chu","year":"2020","journal-title":"Cancer Manag. Res."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"425","DOI":"10.1038\/s41392-021-00828-5","article-title":"Targeting PI3K\/Akt Signal Transduction for Cancer Therapy","volume":"6","author":"He","year":"2021","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"4684","DOI":"10.1038\/s41467-020-18504-7","article-title":"MTOR-Mediated Cancer Drug Resistance Suppresses Autophagy and Generates a Druggable Metabolic Vulnerability","volume":"11","author":"Gremke","year":"2020","journal-title":"Nat. Commun."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"304","DOI":"10.1038\/s41419-020-2467-3","article-title":"The Autophagy-Independent Role of BECN1 in Colorectal Cancer Metastasis through Regulating STAT3 Signaling Pathway Activation","volume":"11","author":"Hu","year":"2020","journal-title":"Cell Death Dis."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1840","DOI":"10.3748\/wjg.v25.i15.1840","article-title":"Predictive and Prognostic Implications of 4E-BP1, Beclin-1, and LC3 for Cetuximab Treatment Combined with Chemotherapy in Advanced Colorectal Cancer with Wild-Type KRAS: Analysis from Real-World Data","volume":"25","author":"Guo","year":"2019","journal-title":"WJG"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1209","DOI":"10.1038\/sj.bjc.6605904","article-title":"Beclin 1 Over- and Underexpression in Colorectal Cancer: Distinct Patterns Relate to Prognosis and Tumour Hypoxia","volume":"103","author":"Koukourakis","year":"2010","journal-title":"Br. J. Cancer"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"961","DOI":"10.1038\/nrc2254","article-title":"Role of Autophagy in Cancer","volume":"7","author":"Mathew","year":"2007","journal-title":"Nat. Rev. Cancer"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"54604","DOI":"10.18632\/oncotarget.17554","article-title":"Expression Analysis of LC3B and P62 Indicates Intact Activated Autophagy Is Associated with an Unfavorable Prognosis in Colon Cancer","volume":"8","author":"Niklaus","year":"2017","journal-title":"Oncotarget"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"e1284720","DOI":"10.1080\/2162402X.2017.1284720","article-title":"Tumor SQSTM1 (P62) Expression and T Cells in Colorectal Cancer","volume":"6","author":"Kosumi","year":"2017","journal-title":"OncoImmunology"},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Ho, W.Y., Yeap, S.K., Ho, C.L., Rahim, R.A., and Alitheen, N.B. (2012). Development of Multicellular Tumor Spheroid (MCTS) Culture from Breast Cancer Cell and a High Throughput Screening Method Using the MTT Assay. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0044640"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/24\/16098\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:43:12Z","timestamp":1760146992000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/24\/16098"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,12,17]]},"references-count":42,"journal-issue":{"issue":"24","published-online":{"date-parts":[[2022,12]]}},"alternative-id":["ijms232416098"],"URL":"https:\/\/doi.org\/10.3390\/ijms232416098","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,12,17]]}}}