Selective ascorbate toxicity in malignant mesothelioma a redox trojan mechanism

We studied the mechanism of ascorbate toxicity in malignant mesothelioma (MMe) cells. Neutral red uptake showed that ascorbate, but not dehydroascorbate, was highly toxic in the MMe cell lines REN and MM98, and less toxic in immortalized (human mesothelial cells-htert) and primary mesothelial cells. Ascorbate transport inhibitors phloretin, sodium azide, and ouabain d id notreduce ascorbate toxicity. Ascorbate promoted the formation of H ₂O₂ in the cell medium, and its toxicity was suppressed by extracellular catalase, but the concentration of endogenous catalase washigher inMMecells than in normal cells.The confocalimaging of cells loaded with the dihydrhodamine 123 reactive oxygen species probe showed that ascorbate caused a strong increase of rhodamine fluorescence in MMe cells, but not in mesothelial cells. MMe cells showed a higher production of superoxide and NADPH oxidase (NOX)4 expression than did mesothelial cells. Two inhibitors of cellular superoxide sources (apocynin and rotenone) reduced ascorbate toxicity and the ascorbate-induced rise in rhodamine fluorescence. NOX4small interferingRNAalso reduced ascorbate toxicity in REN cells. Taken together, the data indicate that ascorbate-induced extracellular H₂O₂ production induces a strong oxidative stress in MMe cells because of their high rate of superoxide production. This explains the selective toxicity of ascorbate in MMe cells, and suggests its possible use in the clinical treatment of malignant mesothelioma.