title: Copper diethyldithiocarbamate as an activator of Nrf2 in cultured vascular endothelial cells.
abstract: The interest in organic-inorganic hybrid molecules as molecular probes for biological systems has been growing rapidly. Such hybrid molecules exhibit unique biological activities. Herein, copper(II) bis(diethyldithiocarbamate) (Cu10) was found to activate the transcription factor NF-E2-related factor 2 (Nrf2), which is responsible for regulating antioxidant and phase II xenobiotic enzymes, in vascular endothelial cells. The copper complex rapidly accumulated within cells and induced nuclear translocation of Nrf2, leading to upregulation of the expression of downstream proteins without cytotoxic effects. However, while copper bis(2-hydroxyethyl)dithiocarbamate activated Nrf2, copper ion, diethyldithiocarbamate ligand with or without zinc or iron failed to exhibit this activity. Intracellular accumulation of Cu10 was higher than that of Cu(II) and Cu(I). While the accumulation of copper(II) bis(dimethyldithiocarbamate) was reduced by small interfering RNA (siRNA)-mediated knockdown of the copper transporter CTR1, the knockdown did not affect Cu10 accumulation, indicating that Cu10 rapidly enters vascular endothelial cells via CTR1-independent mechanisms. In addition, copper and iron complexes with other ligands tested could not activate Nrf2, suggesting that the intramolecular interaction between copper and dithiocarbamate ligand is important for the activation of the transcription factor. Cu10 induced the expression of heme oxygenase-1, NAD(P)H quinone oxidoreductase 1, and γ-glutamylcysteine synthetase, downstream proteins of Nrf2. It was suggested that Cu10-induced activation of Nrf2 was due to proteasome inhibition as well as binding to Kelch-like ECH-associated protein 1. Since the effects of Cu10 on vascular endothelial cells are unique and diverse, the copper complex may be a good molecular probe to analyze the functions of the cells.
authors.affiliation: Department of Environmental Health, Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki, Noda, 278-8510, Japan.
authors.forename: Tomoya
authors.initials: T
authors.lastname: Fujie
authors.affiliation: Department of Environmental Health, Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki, Noda, 278-8510, Japan.
authors.forename: Masaki
authors.initials: M
authors.lastname: Murakami
authors.affiliation: Department of Environmental Health, Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki, Noda, 278-8510, Japan.
authors.forename: Eiko
authors.initials: E
authors.lastname: Yoshida
authors.affiliation: Graduate School of Science and Research Center for Materials Science, Nagoya University, Chikusa, Nagoya, 464-8602, Japan.
authors.forename: Tadashi
authors.initials: T
authors.lastname: Tachinami
authors.affiliation: Environmental Biology Laboratory, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, 305-8575, Japan.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Shinkai
authors.affiliation: Department of Environmental Health, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, 192-0392, Japan.
authors.forename: Yasuyuki
authors.initials: Y
authors.lastname: Fujiwara
authors.affiliation: Department of Environmental Health, Faculty of Pharmaceutical Sciences, Toho University, 2-2-1 Miyama, Funabashi, 274-8510, Japan.
authors.forename: Chika
authors.initials: C
authors.lastname: Yamamoto
authors.affiliation: Environmental Biology Laboratory, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, 305-8575, Japan.
authors.forename: Yoshito
authors.initials: Y
authors.lastname: Kumagai
authors.affiliation: Graduate School of Science and Research Center for Materials Science, Nagoya University, Chikusa, Nagoya, 464-8602, Japan. h_naka@nagoya-u.jp.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Naka
authors.affiliation: Department of Environmental Health, Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki, Noda, 278-8510, Japan. t-kaji@rs.tus.ac.jp.
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Kaji
mesh_terms: D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D003300:Copper / Q000737:chemistry*; D004050:Ditiocarb / Q000737:chemistry*; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D008024:Ligands; D051267:NF-E2-Related Factor 2 / Q000378:metabolism*
chemical_list: D008024:Ligands; D051267:NF-E2-Related Factor 2; D003300:Copper; D004050:Ditiocarb
keywords: Bio-organometallics; Copper(II) bis(diethyldithiocarbamate); Endothelial cell; Keap1; Nrf2; Proteasome
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