Effect of Diquat on the Antioxidant System and Cell Growth in Human Neuroblastoma Cells

• Published in: Toxicology and applied pharmacology Vol. 178; no. 2; pp. 63 - 70
• Main Authors: Slaughter, Mark R, Thakkar, Hansa, O'Brien, Peter J
• Format: Journal Article
• Language: English
• Published: San Diego, CA Elsevier Inc 15.01.2002; Elsevier
• Subjects: antioxidant system; Antioxidants - metabolism; Biological and medical sciences; Brain Neoplasms - enzymology; Brain Neoplasms - metabolism; Brain Neoplasms - pathology; Cell Division - drug effects; cell growth; Cell Survival - drug effects; diquat; Diquat - toxicity; Glutathione - metabolism; Herbicides - toxicity; Humans; Indicators and Reagents; Medical sciences; mitochondria; Mitochondria - drug effects; Mitochondria - enzymology; Neoplasm Proteins - biosynthesis; Neuroblastoma - enzymology; Neuroblastoma - metabolism; Neuroblastoma - pathology; Neurons - drug effects; Neurons - metabolism; Oxidation-Reduction; Oxidative Stress; Pesticides, fertilizers and other agrochemicals toxicology; SH-SY5Y cells; Tetrazolium Salts; Thiazoles; Toxicology; Tumor Cells, Cultured; cell growth; antioxidant system; diquat; SH-SY5Y cells; mitochondria; oxidative stress; Quaternary ammonium compound; Human; Cell proliferation; Oxidative stress; Mitochondria; Toxicity; Pesticides; Diquat; Mechanism of action; In vitro; Herbicide; Tumor cell
• ISSN: 0041-008X; 1096-0333
• DOI: 10.1006/taap.2001.9322

Oxidative stress elicits an adaptive antioxidant response, which varies with tissue type. Diquat, a potent redox cycler that generates reactive oxygen species, has been used to study oxidative stress; however, its effect on the antioxidant system has not been characterized in neuronal cells. Accordingly, we measured antioxidant parameters and cell growth in human neuroblastoma SH-SY5Y cells cultured for 48 h in medium containing 5, 10, or 25 μM diquat dibromide or phosphate-buffered saline. Viable cells were assayed for glutathione (GSH) and activities of catalase (CAT), superoxide dismutase (SOD), glutathione reductase (GR), glutathione peroxidase (GPX), and glucose-6-phosphate dehydrogenase (GPDH). Mitochondrial function was evaluated by glutamate dehydrogenase (GDH) activity and MTT reduction. Diquat caused a marked concentration-related decrease in viable cell count ( by 26, 51, and 87% at 5, 10, and 25 μM diquat). Cell viability was only affected at 10 and 25 μM diquat and did not fully account for the decreased viable cell count. Concentration-related increases also occurred with GSH levels and a majority of antioxidant enzymes activities; however, the mode and magnitude varied with parameter. Increases in GSH, CAT, SOD, and GR were maximal at 25 μM diquat (to 3-, 6-, 2-, and 1.5-fold control values, respectively). GPDH activity was maximal at 10 μM diquat and then decreased to 86% of control activity at 25 μM diquat. GPX activity showed a concentration-related decrease (to 35% of control). Activity of the mitochondrial enzyme GDH increased 3-fold at 25 μM diquat, along with a lesser increase in MTT reduction. We conclude that diquat reduces cell growth in neuroblastoma cells and induces an adaptive antioxidant response, which are concentration dependent and occur at sublethal concentrations. At higher concentrations, diquat alters mitochondrial function and becomes increasingly toxic.