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Two key 8HQs, CQ (5-chloro-7-iodo-8-hydroxyquinoline) and PBT2 (2-(dimethylamino)methyl-5,7-dichloro-8-hydroxyquinoline), have drawn considerable interest and have been the focus of many studies investigating their in vivo properties. These drugs have been described as copper and zinc ionophores because they do not cause metal depletion, as would be expected for a chelation mechanism, but rather cellular accumulation of these ions. In studies of their anti-cancer properties, CQ has been proposed to elicit toxic intracellular copper accumulation and to trigger apoptotic cancer cell death through several possible pathways. In this study we used synchrotron X-ray fluorescence imaging, in combination with biochemical assays and light microscopy, to investigate 8HQ-induced alterations to metal ion homeostasis, as well as cytotoxicity and cell death. We used the bromine fluorescence from a bromine labelled CQ congener (5,7-dibromo-8-hydroxyquinoline; B2Q) to trace the intracellular localization of B2Q following treatment and found that B2Q crosses the cell membrane. We also found that 8HQ co-treatment with Cu(ii) results in significantly increased intracellular copper and significant cytotoxicity compared with 8HQ treatments alone. PBT2 was found to be more cytotoxic, but a weaker Cu(ii) ionophore than other 8HQs. Moreover, treatment of cells with copper in the presence of CQ or B2Q resulted in copper accumulation in the nuclei, while PBT2-guided copper was distributed near to the cell membrane. These results suggest that PBT2 may be acting through a different mechanism than that of other 8HQs to cause the observed cytotoxicity.<\/jats:p>","DOI":"10.1039\/d0mt00222d","type":"journal-article","created":{"date-parts":[[2020,11,2]],"date-time":"2020-11-02T16:46:23Z","timestamp":1604335583000},"page":"1979-1994","update-policy":"https:\/\/doi.org\/10.1039\/rsc_crossmark_policy","source":"Crossref","is-referenced-by-count":23,"title":["PBT2 acts through a different mechanism of action than other 8-hydroxyquinolines: an X-ray fluorescence imaging study"],"prefix":"10.1093","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9185-5052","authenticated-orcid":false,"given":"Kelly L","family":"Summers","sequence":"first","affiliation":[{"name":"Molecular and Environmental Sciences Group, Department of Geological Sciences, College of Arts and Science, University of Saskatchewan, 114 Science Place, Saskatoon, Saskatchewan S7N 5E2, Canada"},{"name":"Department of Chemistry, College of Arts and Science, University of Saskatchewan, 110 Science Place, Saskatoon, Saskatchewan S7N 5C9, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9951-6634","authenticated-orcid":false,"given":"Natalia V","family":"Dolgova","sequence":"additional","affiliation":[{"name":"Molecular and Environmental Sciences Group, Department of Geological Sciences, College of Arts and Science, University of Saskatchewan, 114 Science Place, Saskatoon, Saskatchewan S7N 5E2, Canada"},{"name":"Department of Biochemistry, Microbiology and Immunology, College of Medicine, University of Saskatchewan, 107 Wiggins Road, Saskatoon, Saskatchewan S7N 5E5, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kenneth B","family":"Gagnon","sequence":"additional","affiliation":[{"name":"Department of Anatomy, Physiology and Pharmacology, College of Medicine, University of Saskatchewan, 107 Wiggins Road, Saskatoon, Saskatchewan S7N 5E5, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"George J","family":"Sopasis","sequence":"additional","affiliation":[{"name":"Department of Chemistry, University of Adelaide, South Australia 5005, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ashley K","family":"James","sequence":"additional","affiliation":[{"name":"Molecular and Environmental Sciences Group, Department of Geological Sciences, College of Arts and Science, University of Saskatchewan, 114 Science Place, Saskatoon, Saskatchewan S7N 5E2, Canada"},{"name":"Toxicology Centre, University of Saskatchewan, 44 Campus Drive, Saskatoon, Saskatchewan S7N 5B3, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6061-7155","authenticated-orcid":false,"given":"Barry","family":"Lai","sequence":"additional","affiliation":[{"name":"The Advanced Photon Source, Argonne National Laboratory, 9700 Cass Avenue, Lemont, Illinois 60439, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicole J","family":"Sylvain","sequence":"additional","affiliation":[{"name":"Canadian Light Source Inc., 44 Innovation Blvd, Saskatoon, Saskatchewan S7N 2V3, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3472-8628","authenticated-orcid":false,"given":"Hugh H","family":"Harris","sequence":"additional","affiliation":[{"name":"Department of Chemistry, University of Adelaide, South Australia 5005, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Helen K","family":"Nichol","sequence":"additional","affiliation":[{"name":"Department of Anatomy, Physiology and Pharmacology, College of Medicine, University of Saskatchewan, 107 Wiggins Road, Saskatoon, Saskatchewan S7N 5E5, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0420-7493","authenticated-orcid":false,"given":"Graham 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Science, University of Saskatchewan, 114 Science Place, Saskatoon, Saskatchewan S7N 5E2, Canada"},{"name":"Department of Chemistry, College of Arts and Science, University of Saskatchewan, 110 Science Place, Saskatoon, Saskatchewan S7N 5C9, Canada"},{"name":"Department of Chemistry, University of Adelaide, South Australia 5005, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2020,11,10]]},"reference":[{"key":"2021121004462733100_cit1","volume-title":"Derivatives of oxine","author":"Hollingshead","year":"1956"},{"key":"2021121004462733100_cit2","doi-asserted-by":"crossref","first-page":"622","DOI":"10.1212\/01.wnl.0000324620.86645.c1","volume":"71","author":"Schaumburg","year":"2008","journal-title":"Neurology"},{"key":"2021121004462733100_cit3","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/0009-8981(73)90052-1","volume":"47","author":"Tamura","year":"1973","journal-title":"Clin. 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