{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,8]],"date-time":"2025-11-08T17:13:15Z","timestamp":1762621995389,"version":"build-2065373602"},"reference-count":48,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2018,11,11]],"date-time":"2018-11-11T00:00:00Z","timestamp":1541894400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["UID\/Multi\/04349\/2013"],"award-info":[{"award-number":["UID\/Multi\/04349\/2013"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>64CuCl2 has recently been proposed as a promising agent for prostate cancer (PCa) theranostics, based on preclinical studies in cellular and animal models, and on the increasing number of human studies documenting its use for PCa diagnosis. Nevertheless, the use of 64CuCl2 raises important radiobiological questions that have yet to be addressed. In this work, using a panel of PCa cell lines in comparison with a non-tumoral prostate cell line, we combined cytogenetic approaches with radiocytotoxicity assays to obtain significant insights into the cellular consequences of exposure to 64CuCl2. PCa cells were found to exhibit increased 64CuCl2 uptake, which could not be attributed to increased expression of the main copper cellular importer, hCtr1, as had been previously suggested. Early DNA damage and genomic instability were also higher in PCa cells, with the tumoral cell lines exhibiting deficient DNA-damage repair upon exposure to 64CuCl2. This was corroborated by the observation that 64CuCl2 was more cytotoxic in PCa cells than in non-tumoral cells. Overall, we showed for the first time that PCa cells had a higher sensitivity to 64CuCl2 than healthy cells, supporting the idea that this compound deserved to be further evaluated as a theranostic agent in PCa.<\/jats:p>","DOI":"10.3390\/molecules23112944","type":"journal-article","created":{"date-parts":[[2018,11,14]],"date-time":"2018-11-14T02:42:41Z","timestamp":1542163361000},"page":"2944","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":21,"title":["Radiobiological Characterization of 64CuCl2 as a Simple Tool for Prostate Cancer Theranostics"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1960-603X","authenticated-orcid":false,"given":"Joana","family":"Guerreiro","sequence":"first","affiliation":[{"name":"Centro de Ci\u00eancias e Tecnologias Nucleares, Instituto Superior T\u00e9cnico, Universidade de Lisboa, Estrada Nacional 10 (km 139,7), 2695-066 Bobadela LRS, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4197-1754","authenticated-orcid":false,"given":"V\u00edtor","family":"Alves","sequence":"additional","affiliation":[{"name":"Instituto de Ci\u00eancias Nucleares Aplicadas \u00e0 Sa\u00fade, Universidade de Coimbra, P\u00f3lo de Ci\u00eancias da Sa\u00fade, Az. Sta. Comba, 3000-548 Coimbra, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4145-854X","authenticated-orcid":false,"given":"Antero","family":"Abrunhosa","sequence":"additional","affiliation":[{"name":"Instituto de Ci\u00eancias Nucleares Aplicadas \u00e0 Sa\u00fade, Universidade de Coimbra, P\u00f3lo de Ci\u00eancias da Sa\u00fade, Az. Sta. Comba, 3000-548 Coimbra, Portugal"}]},{"given":"Ant\u00f3nio","family":"Paulo","sequence":"additional","affiliation":[{"name":"Centro de Ci\u00eancias e Tecnologias Nucleares, Instituto Superior T\u00e9cnico, Universidade de Lisboa, Estrada Nacional 10 (km 139,7), 2695-066 Bobadela LRS, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0366-8124","authenticated-orcid":false,"given":"Oct\u00e1via","family":"Gil","sequence":"additional","affiliation":[{"name":"Centro de Ci\u00eancias e Tecnologias Nucleares, Instituto Superior T\u00e9cnico, Universidade de Lisboa, Estrada Nacional 10 (km 139,7), 2695-066 Bobadela LRS, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0646-1687","authenticated-orcid":false,"given":"Filipa","family":"Mendes","sequence":"additional","affiliation":[{"name":"Centro de Ci\u00eancias e Tecnologias Nucleares, Instituto Superior T\u00e9cnico, Universidade de Lisboa, Estrada Nacional 10 (km 139,7), 2695-066 Bobadela LRS, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2018,11,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"7","DOI":"10.3322\/caac.21442","article-title":"Cancer statistics, 2018","volume":"68","author":"Siegel","year":"2018","journal-title":"CA Cancer J. Clin."},{"key":"ref_2","unstructured":"Eurostat (2018, November 11). Cancer Statistics\u2013Specific Cancers. Available online: https:\/\/ec.europa.eu\/eurostat\/statistics-explained\/index.php\/Cancer_statistics_-_specific_cancers."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Sumanasuriya, S., and De Bono, J. (2018). Treatment of advanced prostate cancer\u2014A review of current therapies and future promise. Cold Spring Harb. Perspect. Med., 8.","DOI":"10.1101\/cshperspect.a030635"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"470","DOI":"10.1007\/s11604-016-0553-3","article-title":"Update on advances in molecular PET in urological oncology","volume":"34","author":"Kitajima","year":"2016","journal-title":"Jpn J. Radiol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"134","DOI":"10.1016\/j.urolonc.2015.05.015","article-title":"State of the art imaging of prostate cancer","volume":"34","author":"Marko","year":"2016","journal-title":"Urol. Oncol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1455","DOI":"10.2214\/AJR.09.2579","article-title":"Challenges in clinical prostate cancer: Role of imaging","volume":"192","author":"Kelloff","year":"2009","journal-title":"AJR Am. J. Roentgenol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"355","DOI":"10.1126\/scitranslmed.aaf3936","article-title":"Imaging approaches to optimize molecular therapies","volume":"8","author":"Weissleder","year":"2016","journal-title":"Sci. Transl. Med."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Niccoli Asabella, A., Cascini, G.L., Altini, C., Paparella, D., Notaristefano, A., and Rubini, G. (2014). The copper radioisotopes: A systematic review with special interest to 64Cu. Biomed. Res. Int., 2014.","DOI":"10.1155\/2014\/786463"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"4720","DOI":"10.1039\/c3cc41554f","article-title":"Tumour targeting with radiometals for diagnosis and therapy","volume":"49","author":"Ramogida","year":"2013","journal-title":"Chem. Commun. (Camb)"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"4380","DOI":"10.1074\/jbc.M104728200","article-title":"Biochemical characterization of the human copper transporter Ctr1","volume":"277","author":"Lee","year":"2002","journal-title":"J. Biol. Chem."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1103","DOI":"10.1016\/S1359-6446(05)03541-5","article-title":"Anticopper therapy against cancer and diseases of inflammation and fibrosis","volume":"10","author":"Brewer","year":"2005","journal-title":"Drug. Discov. Today"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"19507","DOI":"10.1073\/pnas.1318431110","article-title":"Bioavailable copper modulates oxidative phosphorylation and growth of tumors","volume":"110","author":"Ishida","year":"2013","journal-title":"Proc. Natl. Acad. Sc.i USA"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"117","DOI":"10.2174\/18744710113069990020","article-title":"New issues for copper-64: From precursor to innovative PET tracers in clinical oncology","volume":"6","author":"Evangelista","year":"2013","journal-title":"Curr. Radiopharm."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"784","DOI":"10.1093\/jrr\/rrv042","article-title":"Validation of 64Cu-ATSM damaging DNA via high-LET Auger electron emission","volume":"56","author":"McMillan","year":"2015","journal-title":"J. Radiat. Res."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"330","DOI":"10.1007\/s00259-009-1305-8","article-title":"Radiobiological effects of hypoxia-dependent uptake of 64Cu-ATSM: Enhanced DNA damage and cytotoxicity in hypoxic cells","volume":"37","author":"Weeks","year":"2010","journal-title":"Eur. J. Nucl. Med. Mol. Imaging"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1016\/j.nucmedbio.2004.08.012","article-title":"Basic characterization of 64Cu-ATSM as a radiotherapy agent","volume":"32","author":"Obata","year":"2005","journal-title":"Nucl. Med. Biol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"812","DOI":"10.2967\/jnumed.113.133850","article-title":"Theranostics of malignant melanoma with 64CuCl2","volume":"55","author":"Qin","year":"2014","journal-title":"J. Nucl. Med."},{"key":"ref_18","first-page":"1649","article-title":"PET of human prostate cancer xenografts in mice with increased uptake of 64CuCl2","volume":"47","author":"Peng","year":"2006","journal-title":"J. Nucl. Med."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Ferrari, C., Asabella, A.N., Villano, C., Giacobbi, B., Coccetti, D., Panichelli, P., and Rubini, G. (2015). Copper-64 Dichloride as Theranostic Agent for Glioblastoma Multiforme: A Preclinical Study. Biomed. Res. Int., 2015.","DOI":"10.1155\/2015\/129764"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"108","DOI":"10.1159\/000480328","article-title":"The SGK1 Kinase Inhibitor SI113 Sensitizes Theranostic Effects of the 64CuCl2 in Human Glioblastoma Multiforme Cells","volume":"43","author":"Catalogna","year":"2017","journal-title":"Cell. Physiol. Biochem."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1692","DOI":"10.2967\/jnumed.114.141127","article-title":"Detection of increased 64Cu uptake by human copper transporter 1 gene overexpression using PET with 64CuCl2 in human breast cancer xenograft model","volume":"55","author":"Kim","year":"2014","journal-title":"J. Nucl. Med."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"444","DOI":"10.2967\/jnumed.117.195628","article-title":"64CuCl2 PET\/CT in Prostate Cancer Relapse","volume":"59","author":"Piccardo","year":"2018","journal-title":"J. Nucl. Med."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"482","DOI":"10.1007\/s12149-015-0968-4","article-title":"Role of 64CuCl2 PET\/CT in staging of prostate cancer","volume":"29","author":"Capasso","year":"2015","journal-title":"Ann. Nucl. Med."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"42544","DOI":"10.1038\/srep42544","article-title":"Evaluation of Acridine Orange Derivatives as DNA-Targeted Radiopharmaceuticals for Auger Therapy: Influence of the Radionuclide and Distance to DNA","volume":"7","author":"Pereira","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"263","DOI":"10.2174\/157016310793360657","article-title":"Targeting the nucleus: An overview of Auger-electron radionuclide therapy","volume":"7","author":"Cornelissen","year":"2010","journal-title":"Curr. Drug Discov. Technol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"622","DOI":"10.2967\/jnumed.113.126979","article-title":"Reduced 64Cu uptake and tumor growth inhibition by knockdown of human copper transporter 1 in xenograft mouse model of prostate cancer","volume":"55","author":"Cai","year":"2014","journal-title":"J. Nucl. Med."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"5858","DOI":"10.1074\/jbc.273.10.5858","article-title":"DNA double-stranded breaks induce histone H2AX phosphorylation on serine 139","volume":"273","author":"Rogakou","year":"1998","journal-title":"J. Biol. Chem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"518","DOI":"10.1667\/RR3431.1","article-title":"Immunofluorescence detection of clustered gamma-H2AX foci induced by HZE-particle radiation","volume":"164","author":"Desai","year":"2005","journal-title":"Rad. Res."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1084","DOI":"10.1038\/nprot.2007.77","article-title":"Cytokinesis-block micronucleus cytome assay","volume":"2","author":"Fenech","year":"2007","journal-title":"Nat. Prot."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2315","DOI":"10.1038\/nprot.2006.339","article-title":"Clonogenic assay of cells in vitro","volume":"1","author":"Franken","year":"2006","journal-title":"Nat. Prot."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1186\/s13550-018-0373-9","article-title":"Biokinetic and dosimetric aspects of 64CuCl2 in human prostate cancer: Possible theranostic implications","volume":"8","author":"Righi","year":"2018","journal-title":"EJNMMI Res."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Cunningham, D., and You, Z. (2015). In vitro and in vivo model systems used in prostate cancer research. J. Biol. Met., 2.","DOI":"10.14440\/jbm.2015.63"},{"key":"ref_33","first-page":"S18","article-title":"18F-Choline, 11C-choline and 11C-acetate PET\/CT: Comparative analysis for imaging prostate cancer patients","volume":"40","author":"Brogsitter","year":"2013","journal-title":"EJNMMI"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"345","DOI":"10.1016\/j.nucmedbio.2013.01.002","article-title":"High tumor uptake of 64Cu: Implications for molecular imaging of tumor characteristics with copper-based PET tracers","volume":"40","author":"Jorgensen","year":"2013","journal-title":"Nucl. Med. Biol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1041","DOI":"10.1369\/jhc.6A6970.2006","article-title":"Expression of the human copper influx transporter 1 in normal and malignant human tissues","volume":"54","author":"Holzer","year":"2006","journal-title":"J. Histochem. Cytochem."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1016\/j.freeradbiomed.2016.12.008","article-title":"Radiation\u2013induced clustered DNA lesions: Repair and mutagenesis","volume":"107","author":"Sage","year":"2017","journal-title":"Free Radic. Biol. Med."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1353","DOI":"10.2967\/jnumed.108.051805","article-title":"Relationship between induction of phosphorylated H2AX and survival in breast cancer cells exposed to 111In-DTPA-hEGF","volume":"49","author":"Cai","year":"2008","journal-title":"J. Nucl. Med."},{"key":"ref_38","first-page":"3369","article-title":"P53 is mutated in a subset of advanced-stage prostate cancers","volume":"53","author":"Bookstein","year":"1993","journal-title":"Cancer Res."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"971","DOI":"10.1016\/S0090-4295(96)00365-2","article-title":"The role of the p53 tumor suppressor gene in prostate cancer: A possible biomarker?","volume":"48","author":"Heidenberg","year":"1996","journal-title":"Urology"},{"key":"ref_40","first-page":"7190","article-title":"Functional p53 increases prostate cancer cell survival after exposure to fractionated doses of ionizing radiation","volume":"63","author":"Scott","year":"2003","journal-title":"Cancer Res."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"2175","DOI":"10.3892\/or.2013.2364","article-title":"Decay of gamma-H2AX foci correlates with potentially lethal damage repair in prostate cancer cells","volume":"29","author":"Hovingh","year":"2013","journal-title":"Oncol. Rep-"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1215","DOI":"10.1016\/j.cell.2015.05.001","article-title":"Integrative clinical genomics of advanced prostate cancer","volume":"161","author":"Robinson","year":"2015","journal-title":"Cell"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"940","DOI":"10.1634\/theoncologist.2016-0135","article-title":"DNA Repair Deficiency Is Common in Advanced Prostate Cancer: New Therapeutic Opportunities","volume":"21","author":"Dhawan","year":"2016","journal-title":"Oncologist"},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Cheng, H.H. (2018). The resounding effect of DNA repair deficiency in prostate cancer. Urol. Oncol.","DOI":"10.1016\/j.urolonc.2018.02.014"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"98","DOI":"10.1186\/s13550-017-0346-4","article-title":"Biodistribution and radiation dosimetry of [(64)Cu]copper dichloride: First in human study in healthy volunteers","volume":"7","author":"Rios","year":"2017","journal-title":"EJNMMI Res."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1740013","DOI":"10.1142\/S0217732317400132","article-title":"Production of copper\u201364 and gallium\u201368 with a medical cyclotron using liquid targets","volume":"32","author":"Alves","year":"2017","journal-title":"Mod. Phys. Lett. A"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1016\/j.jinorgbio.2016.11.026","article-title":"Biophysical characterization and antineoplastic activity of new bis(thiosemicarbazonato) Cu(II) complexes","volume":"167","author":"Palma","year":"2017","journal-title":"J. Inorg. Biochem."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"431","DOI":"10.2203\/dose-response.12-036.Belchior","article-title":"Dose and Time Dependence of Targeted and Untargeted Effects after Very Low Doses of Alpha-Particle Irradiation of Human Lung Cancer Cells","volume":"11","author":"Belchior","year":"2013","journal-title":"Dose Response"}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/23\/11\/2944\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T15:29:05Z","timestamp":1760196545000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/23\/11\/2944"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,11,11]]},"references-count":48,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2018,11]]}},"alternative-id":["molecules23112944"],"URL":"https:\/\/doi.org\/10.3390\/molecules23112944","relation":{},"ISSN":["1420-3049"],"issn-type":[{"type":"electronic","value":"1420-3049"}],"subject":[],"published":{"date-parts":[[2018,11,11]]}}}