title: Superoxide anions are involved in doxorubicin-induced ERK activation in hepatocyte cultures.
abstract: Doxorubicin (DOX), an antineoplastic agent widely used for the treatment of cancer, belongs to the anthracycline family of antitumor antibiotics. DOX may undergo one-electron reduction to the corresponding semiquinone free radical by flavin-containing reductases. Under aerobic conditions, the semiquinone radical reacts rapidly with oxygen to generate superoxide anion, undergoing redox cycling. At moderate concentrations, reactive oxygen species (ROS) play an important role as regulatory mediators in signaling processes. We have shown that DOX increased phosphorylation of enzymes comprising mitogen-activated protein (MAP) kinase cascades in primary hepatocyte cultures, and that this action was independent of oxidant damage. In particular, extracellular signal-regulated kinase (ERK) was phosphorylated by the drug treatment. In this work, we have determined the possible involvement of particular free radicals in DOX-induced ERK phosphorylation in hepatocyte cultures by using specific free radical scavengers. The levels of ERK phosphorylation were measured by Western blot analysis with an anti-Thr202/Tyr204-phosphorylated p44/p42 MAPK antibody. Deferoxamine (DFO; iron chelator), catalase (hydrogen peroxide-removing enzyme), or alpha-tocopherol (peroxyl-radical scavenger) did not affect DOX-increased ERK phosphorylation levels. However, the cell-permeable superoxide dismutase mimetic MnTBAP and the flavin-containing enzyme inhibitor diphenyleneiodonium reverted DOX-induced effects. These results suggest that superoxide anions, probably generated by DOX metabolism, are involved in the effects of the anthracycline on the MAP kinase cascade activation.
authors: Department of Physiology, Medicine and Dentistry School, University of the Basque Country, 48080-Bilbao, Spain.
authors: Rosaura
authors: R
authors: Navarro
authors: Idoia
authors: I
authors: Busnadiego
authors: M Begoña
authors: MB
authors: Ruiz-Larrea
authors: José Ignacio
authors: JI
authors: Ruiz-Sanz
mesh_terms: D000818:Animals; D000838:Anions; D000970:Antineoplastic Agents / Q000494:pharmacology*; D015153:Blotting, Western; D004317:Doxorubicin / Q000494:pharmacology*; D004789:Enzyme Activation; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism*; D022781:Hepatocytes / Q000187:drug effects* / Q000201:enzymology; D008297:Male; D010766:Phosphorylation; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000378:metabolism; D013481:Superoxides / Q000378:metabolism*
chemical_list: D000838:Anions; D000970:Antineoplastic Agents; D017382:Reactive Oxygen Species; D013481:Superoxides; D004317:Doxorubicin; D048049:Extracellular Signal-Regulated MAP Kinases