title: The role of p53 in the trafficking of copper-64 to tumor cell nuclei.
abstract: Copper-64 (T(1/2) = 12.7 h; beta(+): 17.8%, beta(-): 41%) has applications in both positron emission tomography (PET) imaging and targeted radiotherapy of cancer. Copper-64 radiopharmaceuticals have shown tumor growth inhibition with a relatively low radiation dose in animal models; however, the mechanism of cytotoxicity has not been fully elucidated. Here, we report an investigation on the potential role of the tumor suppressor protein p53 in trafficking (64)Cu to tumor cell nuclei. Two EGFR expressing human colorectal cell lines (HCT 116 +/+ and HCT 116 -/-) that are positive or negative for p53 expression respectively, were used to compare internalization and nuclear localization of [(64)Cu]copper acetate and of (64)Cu-DOTA-cetuximab, a monoclonal anti-EGFR antibody. [(64)Cu]copper acetate uptake into cells was similar between the two cell lines during a 24 h time course. In contrast, the uptake of [(64)Cu]copper acetate in the nuclei of HCT 116 +/+ cells was significantly higher than in HCT 116 -/- cells (p < 0.0001) at 24 h. There was no difference in receptor binding, receptor-mediated internalization, and efflux of (64)Cu-DOTA-cetuximab between the two HCT 116 cells lines. However, nuclear localization of (64)Cu-DOTA-cetuximab showed increased uptake in the nuclei of HCT 116 +/+ cells as early as 4 h. These data demonstrate that (64)Cu is delivered to tumor cell nuclei in a p53 positive cell line in significantly greater amounts than in p53 negative cells by both non-specific and receptor-mediated uptake mechanisms.
authors.affiliation: Mallinckrodt Institute of Radiology, Washington University School of Medicine, Saint Louis, Missouri 63110, USA.
authors.forename: Martin
authors.initials: M
authors.lastname: Eiblmaier
authors.forename: Laura A
authors.initials: LA
authors.lastname: Meyer
authors.forename: Carolyn J
authors.initials: CJ
authors.lastname: Anderson
mesh_terms: D021581:Active Transport, Cell Nucleus; D000911:Antibodies, Monoclonal / Q000493:pharmacokinetics; D061067:Antibodies, Monoclonal, Humanized; D002467:Cell Nucleus / Q000378:metabolism*; D000068818:Cetuximab; D003301:Copper Radioisotopes / Q000493:pharmacokinetics* / Q000627:therapeutic use; D004249:DNA Damage; D066246:ErbB Receptors / Q000378:metabolism; D045325:HCT116 Cells; D006801:Humans; D019275:Radiopharmaceuticals / Q000493:pharmacokinetics*; D016159:Tumor Suppressor Protein p53 / Q000502:physiology*
chemical_list: D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized; D003301:Copper Radioisotopes; D019275:Radiopharmaceuticals; D016159:Tumor Suppressor Protein p53; D066246:ErbB Receptors; D000068818:Cetuximab