Production of hydrogen peroxide by murine epidermal keratinocytes following treatment with the tumor promoter 12-O-tetradecanoylphorbol-13-acetate

The ability of murine epidermal cells to produce intracellular hydrogen peroxide was analyzed by flow cytometry and the measurement of 2',7'- dichlorofluorescin (DCFH) oxidation. Epidermal cells isolated from acetone-treated CD-1 mice for 24 h were relatively homogeneous in cell size and d...

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Published in:	Cancer research (Chicago, Ill.) Vol. 50; no. 18; pp. 6062 - 6067
Main Authors:	ROBERTSON, F. M, BEAVIS, A. J, OBERYSZYN, T. M, O'CONNELL, S. M, DOKIDOS, A, LASKIN, D. L, LASKIN, J. D, REINERS, J. J
Format:	Journal Article
Language:	English
Published:	Philadelphia, PA American Association for Cancer Research 15-09-1990
Subjects:	Animals Biological and medical sciences Carcinogenesis, carcinogens and anticarcinogens Chemical agents Female Flow Cytometry Fluoresceins - metabolism Hydrogen Peroxide - metabolism Keratinocytes - drug effects Keratinocytes - metabolism Keratinocytes - pathology Medical sciences Mice Oxidation-Reduction Oxygen - metabolism Tetradecanoylphorbol Acetate - pharmacology Tumors Cell culture Flow cytometry Oxygen Tumor promotor Rodentia Epidermis Hydrogen Peroxides Cell subpopulation Free radical Vertebrata Mammalia Investigation method Mouse Animal Production Keratinocyte Skin Localization Release
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Summary:	The ability of murine epidermal cells to produce intracellular hydrogen peroxide was analyzed by flow cytometry and the measurement of 2',7'- dichlorofluorescin (DCFH) oxidation. Epidermal cells isolated from acetone-treated CD-1 mice for 24 h were relatively homogeneous in cell size and density and oxidized low levels of DCFH. However, following 12-O-tetradecanoylphorbol-13-acetate (TPA) treatment of mice (10 micrograms; 24 h), two cytokeratin-positive populations of cells were identified that were heterogeneous with respect to size and density. These two TPA-derived cell populations oxidized levels of DCFH that were time and dose dependent and were between 2- and 10-fold higher than levels of DCFH oxidized by cells isolated from acetone-treated mice. The ability of catalase, the enzyme that detoxifies hydrogen peroxide, to suppress DCFH oxidation to control levels suggested that intracellular hydrogen peroxide was responsible for the enhanced rate of DCFH oxidation in epidermal cells isolated from TPA-treated mice. The ability of mouse epidermal keratinocytes to oxidize DCFH in response to TPA treatment was confirmed using a cloned keratinocyte cell line. These results suggest that specific subpopulations of keratinocytes produce elevated levels of intracellular peroxides following treatment with TPA either in vivo or in culture.
ISSN:	0008-5472 1538-7445