{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,28]],"date-time":"2026-01-28T19:45:44Z","timestamp":1769629544465,"version":"3.49.0"},"reference-count":48,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2021,4,20]],"date-time":"2021-04-20T00:00:00Z","timestamp":1618876800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Nutrients"],"abstract":"<jats:p>Magnesium is an essential nutrient involved in many important processes in living organisms, including protein synthesis, cellular energy production and storage, cell growth and nucleic acid synthesis. In this study, we analysed the effect of magnesium deficiency on the proliferation of SaOS-2 osteosarcoma cells. When quiescent magnesium-starved cells were induced to proliferate by serum addition, the magnesium content was 2\u20133 times lower in cells maintained in a medium without magnesium compared with cells growing in the presence of the ion. Magnesium depletion inhibited cell cycle progression and caused the inhibition of cell proliferation, which was associated with mTOR hypophosphorylation at Serine 2448. In order to map the intracellular magnesium distribution, an analytical approach using synchrotron-based X-ray techniques was applied. When cell growth was stimulated, magnesium was mainly localized near the plasma membrane in cells maintained in a medium without magnesium. In non-proliferating cells growing in the presence of the ion, high concentration areas inside the cell were observed. These results support the role of magnesium in the control of cell proliferation, suggesting that mTOR may represent an important target for the antiproliferative effect of magnesium. Selective control of magnesium availability could be a useful strategy for inhibiting osteosarcoma cell growth.<\/jats:p>","DOI":"10.3390\/nu13041376","type":"journal-article","created":{"date-parts":[[2021,4,20]],"date-time":"2021-04-20T05:35:25Z","timestamp":1618896925000},"page":"1376","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Assessment and Imaging of Intracellular Magnesium in SaOS-2 Osteosarcoma Cells and Its Role in Proliferation"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3874-6815","authenticated-orcid":false,"given":"Concettina","family":"Cappadone","sequence":"first","affiliation":[{"name":"Department of Pharmacy and Biotechnology, University of Bologna, 33, 40127 Bologna, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1077-0098","authenticated-orcid":false,"given":"Emil","family":"Malucelli","sequence":"additional","affiliation":[{"name":"Department of Pharmacy and Biotechnology, University of Bologna, 33, 40127 Bologna, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maddalena","family":"Zini","sequence":"additional","affiliation":[{"name":"Department of Biomedical and Neuromotor Sciences, Alma Mater Studiorum, University of Bologna, 33, 40126 Bologna, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3599-7034","authenticated-orcid":false,"given":"Giovanna","family":"Farruggia","sequence":"additional","affiliation":[{"name":"Department of Pharmacy and Biotechnology, University of Bologna, 33, 40127 Bologna, Italy"},{"name":"INBB\u2014Biostructures and Biosystems National Institute, 00136 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4174-4164","authenticated-orcid":false,"given":"Giovanna","family":"Picone","sequence":"additional","affiliation":[{"name":"Department of Pharmacy and Biotechnology, University of Bologna, 33, 40127 Bologna, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2457-3618","authenticated-orcid":false,"given":"Alessandra","family":"Gianoncelli","sequence":"additional","affiliation":[{"name":"Elettra\u2014Sincrotrone Trieste, 34149 Trieste, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7177-9096","authenticated-orcid":false,"given":"Andrea","family":"Notargiacomo","sequence":"additional","affiliation":[{"name":"Institute for Photonics and Nanotechnologies, Consiglio Nazionale delle Ricerche, 00156 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michela","family":"Fratini","sequence":"additional","affiliation":[{"name":"Institute of Nanotechnology-CNR c\/o Physics Department at \u2018Sapienza\u2019 University, 00185 Rome, Italy"},{"name":"IRCCS Fondazione Santa Lucia, 00179 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1780-6804","authenticated-orcid":false,"given":"Carla","family":"Pignatti","sequence":"additional","affiliation":[{"name":"Department of Biomedical and Neuromotor Sciences, Alma Mater Studiorum, University of Bologna, 33, 40126 Bologna, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1426-6400","authenticated-orcid":false,"given":"Stefano","family":"Iotti","sequence":"additional","affiliation":[{"name":"Department of Pharmacy and Biotechnology, University of Bologna, 33, 40127 Bologna, Italy"},{"name":"INBB\u2014Biostructures and Biosystems National Institute, 00136 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Claudio","family":"Stefanelli","sequence":"additional","affiliation":[{"name":"Department for Life Quality Studies, Alma Mater Studiorum, University of Bologna, 47921 Rimini, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,4,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1152\/physrev.00012.2014","article-title":"Magnesium in Man: Implications for Health and Disease","volume":"95","author":"Hoenderop","year":"2015","journal-title":"Physiol. Rev."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1054\/mehy.2000.1133","article-title":"The Multifaceted and Widespread Pathology of Magnesium Deficiency","volume":"56","author":"Johnson","year":"2001","journal-title":"Med. Hypotheses"},{"key":"ref_3","unstructured":"(2021, March 06). NHANES Questionnaires, Datasets, and Related Documentation, Available online: https:\/\/wwwn.cdc.gov\/nchs\/nhanes\/continuousnhanes\/default.aspx?BeginYear=2005."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"311","DOI":"10.1002\/bies.20183","article-title":"Magnesium: The Missing Element in Molecular Views of Cell Proliferation Control","volume":"27","author":"Rubin","year":"2005","journal-title":"Bioessays"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"164","DOI":"10.1016\/j.bbabio.2005.01.008","article-title":"Quantitative Mathematical Expressions for Accurate in Vivo Assessment of Cytosolic [ADP] and \u0394G of ATP Hydrolysis in the Human Brain and Skeletal Muscle","volume":"1708","author":"Iotti","year":"2005","journal-title":"Biochim. Biophys. Acta (BBA) Bioenerg."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"375","DOI":"10.1038\/nature17407","article-title":"Daily Magnesium Fluxes Regulate Cellular Timekeeping and Energy Balance","volume":"532","author":"Feeney","year":"2016","journal-title":"Nature"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"528","DOI":"10.1097\/00004647-199905000-00007","article-title":"Low Brain Intracellular Free Magnesium in Mitochondrial Cytopathies","volume":"19","author":"Barbiroli","year":"1999","journal-title":"J. Cereb. Blood Flow Metab."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2178","DOI":"10.1093\/jn\/107.12.2178","article-title":"Effects of Magnesium Deficiency on Protein and Nucleic Acid Synthesis in Vivo","volume":"107","author":"Zieve","year":"1977","journal-title":"J. Nutr."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"ec269","DOI":"10.1126\/scisignal.aal3828","article-title":"A Ribosomal Strategy for Magnesium Deficiency","volume":"9","author":"Mushegian","year":"2016","journal-title":"Sci. Signal."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1016\/j.abb.2006.03.026","article-title":"The Logic of the Membrane, Magnesium, Mitosis (MMM) Model for the Regulation of Animal Cell Proliferation","volume":"458","author":"Rubin","year":"2007","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"378","DOI":"10.1016\/j.ctrv.2009.01.003","article-title":"Magnesium and Tumors: Ally or Foe?","volume":"35","author":"Wolf","year":"2009","journal-title":"Cancer Treat. Rev."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1043","DOI":"10.1126\/science.1058519","article-title":"TRP-PLIK, a Bifunctional Protein with Kinase and Ion Channel Activities","volume":"291","author":"Runnels","year":"2001","journal-title":"Science"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Zou, Z.-G., Rios, F.J., Montezano, A.C., and Touyz, R.M. (2019). TRPM7, Magnesium, and Signaling. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20081877"},{"key":"ref_14","first-page":"215","article-title":"Magnesium and Calcium in Drinking Water and the Risk of Death from Esophageal Cancer","volume":"15","author":"Yang","year":"2002","journal-title":"Magnes. Res."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1577","DOI":"10.1002\/ijc.32939","article-title":"Magnesium Intake and Primary Liver Cancer Incidence and Mortality in the Prostate, Lung, Colorectal and Ovarian Cancer Screening Trial","volume":"147","author":"Zhong","year":"2020","journal-title":"Int. J. Cancer"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"417","DOI":"10.1042\/CS20120086","article-title":"Magnesium and Its Transporters in Cancer: A Novel Paradigm in Tumour Development","volume":"123","author":"Wolf","year":"2012","journal-title":"Clin. Sci."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1007\/s12011-018-1583-8","article-title":"The Impact of Trace Minerals on Bone Metabolism","volume":"188","year":"2019","journal-title":"Biol. Trace Elem. Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"105400","DOI":"10.1016\/j.jsbmb.2019.105400","article-title":"Anabolic Effects of Vitamin D and Magnesium in Aging Bone","volume":"193","author":"Erem","year":"2019","journal-title":"J. Steroid Biochem. Mol. Biol."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Mammoli, F., Castiglioni, S., Parenti, S., Cappadone, C., Farruggia, G., Iotti, S., Davalli, P., Maier, J.A.M., Grande, A., and Frassineti, C. (2019). Magnesium Is a Key Regulator of the Balance between Osteoclast and Osteoblast Differentiation in the Presence of Vitamin D3. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20020385"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Picone, G., Cappadone, C., Pasini, A., Lovecchio, J., Cortesi, M., Farruggia, G., Lombardo, M., Gianoncelli, A., Mancini, L., and Ralf, H.M. (2020). Analysis of Intracellular Magnesium and Mineral Depositions during Osteogenic Commitment of 3D Cultured Saos2 Cells. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21072368"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"461","DOI":"10.1038\/nprot.2016.183","article-title":"Synthesis of a Highly Mg2+-Selective Fluorescent Probe and Its Application to Quantifying and Imaging Total Intracellular Magnesium","volume":"12","author":"Sargenti","year":"2017","journal-title":"Nat. Protoc."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"7897","DOI":"10.1021\/jm061077m","article-title":"Synthesis and Antitumor Activity of Guanylhydrazones from 6-(2,4-Dichloro-5-Nitrophenyl)Imidazo[2,1-b]Thiazoles and 6-Pyridylimidazo[2,1-b]Thiazoles","volume":"49","author":"Andreani","year":"2006","journal-title":"J. Med. Chem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1007\/s00216-017-0725-8","article-title":"Single Cell versus Large Population Analysis: Cell Variability in Elemental Intracellular Concentration and Distribution","volume":"410","author":"Malucelli","year":"2018","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"5108","DOI":"10.1021\/ac5008909","article-title":"Quantitative Chemical Imaging of the Intracellular Spatial Distribution of Fundamental Elements and Light Metals in Single Cells","volume":"86","author":"Malucelli","year":"2014","journal-title":"Anal. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1526","DOI":"10.1107\/S1600577516014405","article-title":"Current Status of the TwinMic Beamline at Elettra: A Soft X-Ray Transmission and Emission Microscopy Station","volume":"23","author":"Gianoncelli","year":"2016","journal-title":"J. Synchrotron Radiat."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"182001","DOI":"10.1088\/1742-6596\/425\/18\/182001","article-title":"Recent Developments at the TwinMic Beamline at ELETTRA: An 8 SDD Detector Setup for Low Energy X-Ray Fluorescence","volume":"425","author":"Gianoncelli","year":"2013","journal-title":"J. Phys. Conf. Ser."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"251117","DOI":"10.1063\/1.2422908","article-title":"Scanning Transmission X-Ray Microscopy with a Configurable Detector","volume":"89","author":"Gianoncelli","year":"2006","journal-title":"Appl. Phys. Lett."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1684\/mrh.2020.0464","article-title":"The Assessment of Intracellular Magnesium: Different Strategies to Answer Different Questions","volume":"33","author":"Picone","year":"2020","journal-title":"Magnes. Res."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/S0065-230X(05)93001-7","article-title":"Central Roles of Mg2+ and MgATP2\u2212 in the Regulation of Protein Synthesis and Cell Proliferation: Significance for Neoplastic Transformation","volume":"93","author":"Rubin","year":"2005","journal-title":"Adv. Cancer Res."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"3563","DOI":"10.1002\/jcb.23284","article-title":"Magnesium Deficiency Suppresses Cell Cycle Progression Mediated by Increase in Transcriptional Activity of P21(Cip1) and P27(Kip1) in Renal Epithelial NRK-52E Cells","volume":"112","author":"Ikari","year":"2011","journal-title":"J. Cell Biochem."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1042\/CS20070129","article-title":"Cell (Patho)Physiology of Magnesium","volume":"114","author":"Wolf","year":"2008","journal-title":"Clin. Sci."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Shamloo, B., and Usluer, S. (2019). P21 in Cancer Research. Cancers, 11.","DOI":"10.3390\/cancers11081178"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1007\/s00726-008-0230-7","article-title":"MTOR Phosphorylated at S2448 Binds to Raptor and Rictor","volume":"38","author":"Rosner","year":"2010","journal-title":"Amino Acids"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"683","DOI":"10.1016\/j.semcdb.2010.03.002","article-title":"Physiological Role of Autophagy as an Intracellular Recycling System: With an Emphasis on Nutrient Metabolism","volume":"21","author":"Kuma","year":"2010","journal-title":"Semin. Cell Dev. Biol."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Yoshii, S.R., and Mizushima, N. (2017). Monitoring and Measuring Autophagy. Int. J. Mol. Sci., 18.","DOI":"10.3390\/ijms18091865"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1016\/j.bbrc.2020.12.008","article-title":"Comparison of Chloroquine-like Molecules for Lysosomal Inhibition and Measurement of Autophagic Flux in the Brain","volume":"534","author":"Fourrier","year":"2021","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"8199","DOI":"10.3390\/nu7095388","article-title":"Magnesium in Prevention and Therapy","volume":"7","author":"Schmidt","year":"2015","journal-title":"Nutrients"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"92","DOI":"10.1684\/mrh.2011.0285","article-title":"Magnesium and Cancer: A Dangerous Liason","volume":"24","author":"Castiglioni","year":"2011","journal-title":"Magnes. Res."},{"key":"ref_39","first-page":"49","article-title":"Magnesium in Health and Disease","volume":"13","author":"Romani","year":"2013","journal-title":"Met. Ions Life Sci."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"5221","DOI":"10.1039\/C6AN01091A","article-title":"Where Is It and How Much? Mapping and Quantifying Elements in Single Cells","volume":"141","author":"Malucelli","year":"2016","journal-title":"Analyst"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"313","DOI":"10.1002\/(SICI)1097-4644(19980901)70:3<313::AID-JCB4>3.0.CO;2-Q","article-title":"Magnesium Restriction Induces Granulocytic Differentiation and Expression of P27Kip1 in Human Leukemic HL-60 Cells","volume":"70","author":"Covacci","year":"1998","journal-title":"J. Cell Biochem."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1002\/(SICI)1097-4652(199908)180:2<245::AID-JCP12>3.0.CO;2-R","article-title":"Magnesium Depletion Causes Growth Inhibition, Reduced Expression of Cyclin D1, and Increased Expression of P27KIP1 in Normal but Not in Transformed Mammary Epithelial Cells","volume":"180","author":"Sgambato","year":"1999","journal-title":"J. Cell. Physiol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"829","DOI":"10.1016\/j.cellsig.2012.12.023","article-title":"ERK1\/2-Dependent Activation of MTOR\/MTORC1\/P70S6K Regulates Thrombin-Induced RPE Cell Proliferation","volume":"25","author":"Parrales","year":"2013","journal-title":"Cell Signal."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"309","DOI":"10.1097\/MEG.0b013e32835c073c","article-title":"Nonlinear Association between Magnesium Intake and the Risk of Colorectal Cancer","volume":"25","author":"Qu","year":"2013","journal-title":"Eur. J. Gastroenterol. Hepatol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1499","DOI":"10.1093\/jn\/nxaa040","article-title":"Dietary Supplement Use among Adult Cancer Survivors in the United States","volume":"150","author":"Du","year":"2020","journal-title":"J. Nutr."},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Zak\u0142os-Szyda, M., Nowak, A., Pietrzyk, N., and Pods\u0119dek, A. (2020). Viburnum Opulus L. Juice Phenolic Compounds Influence Osteogenic Differentiation in Human Osteosarcoma Saos-2 Cells. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21144909"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"716","DOI":"10.1111\/j.1600-0501.2010.02046.x","article-title":"Effects of Magnesium Intake Deficiency on Bone Metabolism and Bone Tissue around Osseointegrated Implants: Effects of Magnesium Intake Deficiency on Bone Tissue","volume":"22","author":"Belluci","year":"2011","journal-title":"Clin. Oral Implants Res."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1016\/j.maturitas.2020.05.020","article-title":"Calcium, Vitamin D, Vitamin K2, and Magnesium Supplementation and Skeletal Health","volume":"140","author":"Capozzi","year":"2020","journal-title":"Maturitas"}],"container-title":["Nutrients"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6643\/13\/4\/1376\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T05:50:00Z","timestamp":1760161800000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6643\/13\/4\/1376"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,4,20]]},"references-count":48,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2021,4]]}},"alternative-id":["nu13041376"],"URL":"https:\/\/doi.org\/10.3390\/nu13041376","relation":{},"ISSN":["2072-6643"],"issn-type":[{"value":"2072-6643","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,4,20]]}}}