Stimulation of Hydrogen Peroxide Production by Drinking Water Contaminants in HL-60 Cells Sensitized by Retinoic Acid

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Chemical carcinogens, such as chloroform and trichloroethylene, are present in drinking water in Japan. As these contaminants are believed to have a role in carcinogenesis, we examined if chloroform and trichloroethylene, as well as methylene chloride, xylene, benzene, and ethanol, have the ability to generate hydrogen peroxide (H_2O_2) in human polymorphonuclear leukocytes (PMN) and human leukemia (HL-60) cells. Methylene chloride, benzene, xylene, trichloroethylene, and ethanol did not increase cellular H_2O_2 production as measured by flow cytometry nor as observed by confocal laser microscopy. In PMN and RA-untreated HL-60 cells chloroform did not significantly affect H_2O_2 levels. However, in HL-60 cells sensitized by pretreatment of 10 nM retinoic acid (RA) for 12 h, chloroform induced a significant increase in H_2O_2, but the increase induced by trichloroethylene was not significant. The observed increase in fluorescence was confirmed using a confocal laser microscope. These results indicate that chloroform and trichloroethylene may stimulate H_2O_2 production in HL-60 cells sensitized by pretreatment of RA. Our method may be useful to test if weak stimulants can stimulate intracellular H_2O_2 production.

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