title: Role of oxidative stress in geldanamycin-induced cytotoxicity and disruption of Hsp90 signaling complex.
abstract: Heat shock protein 90 (Hsp90) is a chaperone protein regulating PC-12 cell survival by binding and stabilizing Akt, Raf-1, and Cdc37. Hsp90 inhibitor geldanamycin (GA) cytotoxicity has been attributed to the disruption of Hsp90 binding, and the contribution of oxidative stress generated by its quinone group has not been studied in this context. Reactive oxygen species (ROS) and cell survival were assessed in PC-12 cells exposed to GA or menadione (MEN), and Akt, Raf-1, and Cdc37 expression and binding to Hsp90 were determined. GA disrupted Hsp90 binding and increased ROS production starting at 1 h, and cell death occurred at 6 h, inhibited by N-acetylcysteine (NAC) without preventing dissociation of proteins. At 24 h, NAC prevented cytotoxicity and Hsp90 complex disruption. However, MnTBAP antioxidant treatment failed to inhibit GA cytotoxicity, suggesting that NAC acts by restoring glutathione. In contrast, 24 h MEN treatment induced cytotoxicity without disrupting Hsp90 binding. GA and MEN decreased Hsp90-binding protein expression, and proteasomal inhibition prevented MEN-, but not GA-induced degradation. In conclusion, whereas MEN cytotoxicity is mediated by ROS and proteasomal degradation, GA-induced cytotoxicity requires ROS but induces Hsp90 complex dissociation and proteasome-independent protein degradation. These differences between MEN- and GA-induced cytotoxicity may allow more specific targeting of cancer cells.
authors.affiliation: Department of Pharmacology and Toxicology, University of Louisville, Louisville, KY 40292, USA.
authors.forename: Christina B
authors.initials: CB
authors.lastname: Clark
authors.forename: Madhavi J
authors.initials: MJ
authors.lastname: Rane
authors.forename: Delphine
authors.initials: D
authors.lastname: El Mehdi
authors.forename: Cynthia J
authors.initials: CJ
authors.lastname: Miller
authors.forename: Leroy R
authors.initials: LR
authors.lastname: Sachleben
authors.forename: Evelyne
authors.initials: E
authors.lastname: Gozal
mesh_terms: D000818:Animals; D016227:Benzoquinones / Q000633:toxicity*; D002470:Cell Survival / Q000187:drug effects; D004354:Drug Screening Assays, Antitumor; D018841:HSP90 Heat-Shock Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D047029:Lactams, Macrocyclic / Q000633:toxicity*; D018384:Oxidative Stress / Q000187:drug effects*; D016716:PC12 Cells; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D014407:Tumor Cells, Cultured; D024483:Vitamin K 3 / Q000494:pharmacology
chemical_list: D016227:Benzoquinones; D018841:HSP90 Heat-Shock Proteins; D047029:Lactams, Macrocyclic; D017382:Reactive Oxygen Species; D024483:Vitamin K 3; D046988:Proteasome Endopeptidase Complex; C001277:geldanamycin
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