{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,4]],"date-time":"2025-11-04T10:41:01Z","timestamp":1762252861562,"version":"build-2065373602"},"reference-count":46,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2018,10,28]],"date-time":"2018-10-28T00:00:00Z","timestamp":1540684800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>Hepatoblastoma incidence has been associated with different environmental factors even if no data are reported about a correlation between aflatoxin exposure and hepatoblastoma initiation. Considering that hepatoblastoma develops in infants and children and aflatoxin M1 (AFM1), the aflatoxin B1 (AFB1) hydroxylated metabolite, can be present in mothers\u2019 milk and in marketed milk products, in this study we decided to test the effects of AFM1 on a hepatoblastoma cell line (HepG2). Firstly, we evaluated the effects of AFM1 on the cell viability, apoptosis, cell cycle, and metabolomic and cytokinomic profile of HepG2 cells after treatment. AFM1 induced: (1) a decrease of HepG2 cell viability, reaching IC50 at 9 \u00b5M; (2) the blocking of the cell cycle in the G0\/G1 phase; (3) the decrease of formiate levels and incremented level of some amino acids and metabolites in HepG2 cells after treatment; and (4) the increase of the concentration of three pro-inflammatory cytokines, IL-6, IL-8, and TNF-\u03b1, and the decrease of the anti-inflammatory interleukin, IL-4. Our results show that AFM1 inhibited the growth of HepG2 cells, inducing both a modulation of the lipidic, glycolytic, and amino acid metabolism and an increase of the inflammatory status of these cells.<\/jats:p>","DOI":"10.3390\/toxins10110436","type":"journal-article","created":{"date-parts":[[2018,10,29]],"date-time":"2018-10-29T11:10:41Z","timestamp":1540811441000},"page":"436","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Evaluation of Aflatoxin M1 Effects on the Metabolomic and Cytokinomic Profiling of a Hepatoblastoma Cell Line"],"prefix":"10.3390","volume":"10","author":[{"given":"Silvia","family":"Marchese","sequence":"first","affiliation":[{"name":"Unit\u00e0 di Farmacologia e Tossicologia\u2014Dipartimento di Medicina Veterinaria e Produzioni Animali, Universit\u00e0 degli Studi di Napoli \u201cFederico II\u201d, 80138 Napoli, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Angela","family":"Sorice","sequence":"additional","affiliation":[{"name":"Unit\u00e0 di Farmacologia Sperimentale, Istituto Nazionale Tumori-IRCCS-Fondazione G. Pascale, 80131 Napoli, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andrea","family":"Ariano","sequence":"additional","affiliation":[{"name":"Unit\u00e0 di Farmacologia e Tossicologia\u2014Dipartimento di Medicina Veterinaria e Produzioni Animali, Universit\u00e0 degli Studi di Napoli \u201cFederico II\u201d, 80138 Napoli, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Salvatore","family":"Florio","sequence":"additional","affiliation":[{"name":"Unit\u00e0 di Farmacologia e Tossicologia\u2014Dipartimento di Medicina Veterinaria e Produzioni Animali, Universit\u00e0 degli Studi di Napoli \u201cFederico II\u201d, 80138 Napoli, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alfredo","family":"Budillon","sequence":"additional","affiliation":[{"name":"Unit\u00e0 di Farmacologia Sperimentale, Istituto Nazionale Tumori-IRCCS-Fondazione G. Pascale, 80131 Napoli, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7538-4657","authenticated-orcid":false,"given":"Susan","family":"Costantini","sequence":"additional","affiliation":[{"name":"Unit\u00e0 di Farmacologia Sperimentale, Istituto Nazionale Tumori-IRCCS-Fondazione G. Pascale, 80131 Napoli, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lorella","family":"Severino","sequence":"additional","affiliation":[{"name":"Unit\u00e0 di Farmacologia e Tossicologia\u2014Dipartimento di Medicina Veterinaria e Produzioni Animali, Universit\u00e0 degli Studi di Napoli \u201cFederico II\u201d, 80138 Napoli, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2018,10,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"192","DOI":"10.1053\/j.semdp.2016.12.015","article-title":"Hepatoblastoma","volume":"34","author":"Sharma","year":"2017","journal-title":"Semin. Diagn. Pathol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"843","DOI":"10.1016\/j.humpath.2008.10.022","article-title":"Gene expression profiling reveals signatures characterizing histologic subtypes of hepatoblastoma and global deregulation in cell growth and survival pathways","volume":"40","author":"Adesina","year":"2009","journal-title":"Hum. Pathol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"53","DOI":"10.12688\/f1000research.12239.1","article-title":"Hepatoblastoma: Current understanding, recent advances, and controversies","volume":"7","author":"Czauderna","year":"2018","journal-title":"F1000Research"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"285","DOI":"10.1053\/j.gastro.2005.04.013","article-title":"Epidermal growth factor receptor: A novel target of the Wnt\/beta-catenin pathway in liver","volume":"129","author":"Tan","year":"2005","journal-title":"Gastroenterology"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"812","DOI":"10.1634\/theoncologist.2008-0011","article-title":"Liver tumors in children","volume":"13","author":"Litten","year":"2008","journal-title":"Oncologist"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"e5858","DOI":"10.1097\/MD.0000000000005858","article-title":"Hepatoblastoma in an 11-year-old: Case report and a review of the literature","volume":"96","author":"Pateva","year":"2017","journal-title":"Medicine"},{"key":"ref_7","first-page":"2114s","article-title":"Aflatoxins as Risk Factors for Hepatocellular Carcinoma in Humans","volume":"52","author":"Wogan","year":"1992","journal-title":"Cancer Res."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"152","DOI":"10.1006\/taap.1998.8440","article-title":"Metabolism and toxicity of aflatoxins M1 and B1 in human-derived in vitro systems","volume":"151","author":"Neal","year":"1998","journal-title":"Toxicol. Appl. Pharmacol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"134","DOI":"10.1016\/j.fct.2018.01.036","article-title":"Risk assessment of aflatoxin B1 exposure from maize and peanut consumption in Indonesia using the margin of exposure and liver cancer risk estimation approaches","volume":"113","author":"Nugraha","year":"2018","journal-title":"Food Chem. Toxicol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"128","DOI":"10.1080\/19393210.2017.1282545","article-title":"Aflatoxin M1 in human breast milk in Shahrekord, Iran and association with dietary factors","volume":"10","author":"Jafari","year":"2017","journal-title":"Food Addit. Contam. Part B"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1418","DOI":"10.1016\/j.toxrep.2015.10.011","article-title":"Contamination profile of aflatoxin M1 residues in milk supply chain of Sindh, Pakistan","volume":"2","author":"Jawaid","year":"2015","journal-title":"Toxicol. Rep."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1007\/s12550-016-0265-7","article-title":"Mycotoxins as human carcinogens-the IARC Monographs classification","volume":"33","author":"Ostry","year":"2017","journal-title":"Mycotoxin Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1016\/j.toxicon.2018.04.017","article-title":"Lactoferrin inhibits aflatoxin B1- and aflatoxin M1-induced cytotoxicity and DNA damage in Caco-2, HEK, Hep-G2, and SK-N-SH cells","volume":"150","author":"Zheng","year":"2018","journal-title":"Toxicon"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"16732","DOI":"10.1038\/srep16732","article-title":"miR-34a screened by miRNA profiling negatively regulates Wnt\/\u03b2-catenin signaling pathway in Aflatoxin B1 induced hepatotoxicity","volume":"5","author":"Zhu","year":"2015","journal-title":"Sci. Rep."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1016\/j.toxlet.2015.12.006","article-title":"Crosstalk-eliminated quantitative determination of aflatoxin B1-induced hepatocellular cancer stem cells based on concurrent monitoring of CD133, CD44, and aldehyde dehydrogenase1","volume":"243","author":"Ju","year":"2016","journal-title":"Toxicol. Lett."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"5","DOI":"10.33899\/ijvs.2012.46826","article-title":"Effect of aflatoxin on malondialdehyde, glutathione levels, and stress index in Toxoplasma gondii infected mice","volume":"26","author":"Hassan","year":"2012","journal-title":"Iraqi J. Vet. Sci."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"166","DOI":"10.6026\/97320630005166","article-title":"Human Cytokinome: A new challenge for systems biology","volume":"5","author":"Costantini","year":"2010","journal-title":"Bioinformation"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/s10616-014-9761-9","article-title":"Distinctive pharmacological differences between liver cancer cell lines HepG2 and Hep3B","volume":"67","author":"Qiu","year":"2015","journal-title":"Cytotechnology"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1512","DOI":"10.1016\/j.humpath.2009.07.003","article-title":"HepG2 is a hepatoblastoma-derived cell line","volume":"40","author":"Cheung","year":"2009","journal-title":"Hum. Pathol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1312","DOI":"10.1016\/j.jhep.2014.08.009","article-title":"The genomic landscape of hepatoblastoma and their progenies with HCC-like features","volume":"61","author":"Trippel","year":"2014","journal-title":"J. Hepatol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1210","DOI":"10.1016\/j.fct.2007.01.001","article-title":"A longitudinal assessment of aflatoxin M1 excretion in breast milk of selected Egyptian mothers","volume":"45","author":"Polychronaki","year":"2007","journal-title":"Food Chem. Toxicol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"277","DOI":"10.3920\/WMJ2014.1705","article-title":"Association between aflatoxin M1 exposure through breast milk and growth impairment in infants from Northern Tanzania","volume":"7","author":"Magoha","year":"2014","journal-title":"World Mycotoxin J."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"54","DOI":"10.1016\/j.fct.2015.05.020","article-title":"Aflatoxin B1 and aflatoxin M1 induced cytotoxicity and DNA damage in differentiated and undifferentiated Caco-2 cells","volume":"83","author":"Zhang","year":"2015","journal-title":"Food Chem. Toxicol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1016\/j.fct.2016.07.019","article-title":"Aflatoxin M1 cytotoxicity against human intestinal Caco-2 cells is enhanced in the presence of other mycotoxins","volume":"96","author":"Gao","year":"2016","journal-title":"Food Chem. Toxicol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"353","DOI":"10.3109\/1547691X.2013.848250","article-title":"In vitro study of AFB1 and AFM1 effects on human lymphoblastoid Jurkat T-cell model","volume":"11","author":"Luongo","year":"2014","journal-title":"J. Immunotoxicol."},{"key":"ref_26","first-page":"52","article-title":"Lactate Contribution to the Tumor Microenvironment: Mechanisms, Effects on Immune Cells and Therapeutic Relevance","volume":"7","year":"2016","journal-title":"Front. Immunol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"2550","DOI":"10.1158\/0008-5472.CAN-10-2828","article-title":"Lactate Influx through the Endothelial Cell Monocarboxylate Transporter MCT1 Supports an NF-\u03baB\/IL-8 Pathway that Drives Tumor Angiogenesis","volume":"71","author":"Boidot","year":"2011","journal-title":"Cancer Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"2605","DOI":"10.1021\/pr070063h","article-title":"Metabonomic Studies of Human Hepatocellular Carcinoma Using High-Resolution Magic-Angle Spinning 1H NMR Spectroscopy in Conjunction with Multivariate Data Analysis","volume":"6","author":"Yang","year":"2007","journal-title":"J. Proteome Res."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2077","DOI":"10.1021\/mp1001922","article-title":"Transport and Metabolism of Radiolabeled Choline in Hepatocellular Carcinoma","volume":"7","author":"Kuang","year":"2010","journal-title":"Mol. Pharm."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"262","DOI":"10.3389\/fonc.2016.00262","article-title":"The Tumor Microenvironment Modulates Choline and Lipid Metabolism","volume":"6","author":"Mori","year":"2016","journal-title":"Front. Oncol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"614","DOI":"10.1021\/pr100792q","article-title":"Systems Responses of Rats to Aflatoxin B1 Exposure Revealed with Metabonomic Changes in Multiple Biological Matrices","volume":"10","author":"Zhang","year":"2011","journal-title":"J. Proteome Res."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"474","DOI":"10.1158\/1078-0432.CCR-17-1707","article-title":"Global Metabolic Profiling Identifies a Pivotal Role of Proline and Hydroxyproline Metabolism in Supporting Hypoxic Response in Hepatocellular Carcinoma","volume":"24","author":"Tang","year":"2018","journal-title":"Clin. Cancer Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1016\/j.tibs.2014.02.004","article-title":"Serine and glycine metabolism in cancer","volume":"39","author":"Amelio","year":"2014","journal-title":"Trends Biochem. Sci."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"19","DOI":"10.4062\/biomolther.2017.178","article-title":"Targeting Glutamine Metabolism for Cancer Treatment","volume":"26","author":"Choi","year":"2018","journal-title":"Biomol. Ther."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2499","DOI":"10.1093\/carcin\/bgs303","article-title":"Preventive effects of branched-chain amino acid supplementation on the spontaneous development of hepatic preneoplastic lesions in C57BL\/KsJ-db\/db obese mice","volume":"33","author":"Terakura","year":"2012","journal-title":"Carcinogenesis"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1097\/MCO.0000000000000430","article-title":"Branched-chain amino acid metabolism in cancer","volume":"21","author":"Ananieva","year":"2018","journal-title":"Curr. Opin. Clin. Nutr. Metab. Care"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"360","DOI":"10.3382\/ps.2008-00258","article-title":"Toxicological and gene expression analysis of the impact of aflatoxin B1 on hepatic function of male broiler chicks","volume":"88","author":"Yarru","year":"2009","journal-title":"Poult. Sci."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1093\/toxsci\/kfh016","article-title":"Characteristic expression profiles induced by genotoxic carcinogens in rat liver","volume":"77","author":"Stuart","year":"2004","journal-title":"Toxicol. Sci."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"408","DOI":"10.1016\/j.toxrep.2017.07.006","article-title":"Acute aflatoxin B1-Induced hepatotoxicity alters gene expression and disrupts lipid and lipoprotein metabolism in rats","volume":"4","author":"Rotimi","year":"2017","journal-title":"Toxicol. Rep."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"2471","DOI":"10.3892\/or.2014.3484","article-title":"Expression and roles of fatty acid synthase in hepatocellular carcinoma","volume":"32","author":"Hao","year":"2014","journal-title":"Oncol. Rep."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"362","DOI":"10.1093\/toxsci\/kfg074","article-title":"Immunotoxicity of aflatoxin B1 in rats: Effects on lymphocytes and the inflammatory response in a chronic intermittent dosing study","volume":"73","author":"Hinton","year":"2003","journal-title":"Toxicol. Sci."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"241","DOI":"10.1002\/jat.2866","article-title":"Aflatoxin B1 modulates the expression of phenotypic markers and cytokines by splenic lymphocytes of male F344 rats","volume":"34","author":"Qian","year":"2014","journal-title":"J. Appl. Toxicol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"807","DOI":"10.1093\/intimm\/dxh262","article-title":"Aflatoxin B1 albumin adduct levels and cellular immune status in Ghanaians","volume":"17","author":"Jiang","year":"2005","journal-title":"Int. Immunol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1007\/s11306-018-1330-3","article-title":"Effects of \u03b1-zearalenol on the metabolome of two breast cancer cell lines by 1H-NMR approach","volume":"14","author":"Nittoli","year":"2018","journal-title":"Metabolomics"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"5622","DOI":"10.1038\/s41598-018-24023-9","article-title":"Toxigenic effects of two benthic diatoms upon grazing activity of the sea urchin: Morphological, metabolomic and de novo transcriptomic analysis","volume":"8","author":"Ruocco","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"251","DOI":"10.1093\/nar\/gkv380","article-title":"MetaboAnalyst 3.0\u2014Making metabolomics more meaningful","volume":"43","author":"Xia","year":"2015","journal-title":"Nucleic Acids Res."}],"container-title":["Toxins"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6651\/10\/11\/436\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T15:26:38Z","timestamp":1760196398000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6651\/10\/11\/436"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,10,28]]},"references-count":46,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2018,11]]}},"alternative-id":["toxins10110436"],"URL":"https:\/\/doi.org\/10.3390\/toxins10110436","relation":{},"ISSN":["2072-6651"],"issn-type":[{"type":"electronic","value":"2072-6651"}],"subject":[],"published":{"date-parts":[[2018,10,28]]}}}