Production of Hydrogen Peroxide by Murine Epidermal Keratinocytes following Treatment with the Tumor Promoter 12-O-Tetradecanoylphorbol-13-acetate

Fredika M. Robertson, Andrew J. Beavis, Sean M. O'Connell, Tatiana M. Oberyszyn, Anthea Dokidos, Jeffrey D. Laskin, Debra L. Laskin, John J. Reiners

Research output: Contribution to journalArticlepeer-review

69 Scopus citations

Abstract

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-I mice for 24 h were relatively homogeneous in cell size and density and oxidized low levels of DCFH. However, following 12–0-tetradecanoylphorbol-13-acetate (TPA) treatment of mice (10 μg; 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.

Original languageEnglish (US)
Pages (from-to)6062-6067
Number of pages6
JournalCancer Research
Volume50
Issue number18
StatePublished - Sep 15 1990

All Science Journal Classification (ASJC) codes

  • Oncology
  • Cancer Research

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