TRP-2 expression protects HEK cells from dopamine- and hydroquinone-induced toxicity

Free Radic Biol Med. 2008 Oct 1;45(7):1002-10. doi: 10.1016/j.freeradbiomed.2008.06.030. Epub 2008 Jul 10.

Abstract

We previously reported that melanogenic enzyme TRP-2 (or DCT for DOPAchrome tautomerase) expression in WM35 melanoma cells resulted in increased intracellular GSH levels, reduction in DNA damage induced by free radicals, and decreased cell sensitivity to oxidative stress. These effects seemed to depend on a particular cellular context, because none of them were found to occur in HEK epithelial cells. We postulated that the TRP-2 beneficial effect observed in WM35 cells in the oxidative stress situation may relate to quinone metabolization and, more precisely, to the ability of TRP-2 to clear off related toxic metabolites, resulting in a global redox status modification. Here, a comparative protein expression profiling of catecholamine biosynthesis enzymes and detoxification enzymes was conducted in WM35 melanoma cells and in HEK epithelial cells, in comparison with normal human melanocytes. Results showed that WM35 cells, but not HEK cells, expressed enzymes involved in catecholamine biosynthesis, suggesting that their quinone-related toxic metabolites were present in WM35 cells but not in HEK cells. To address the issue of a possible TRP-2 beneficial effect toward quinone toxicity, cell survival experiments were then conducted in HEK cells using dopamine and hydroquinone at toxic concentrations. We showed that TRP-2 expression significantly reduced HEK cell sensitivity to both compounds. This beneficial property of TRP-2 was likely to depend on the integrity of its DOPAchrome tautomerase catalytic site, because both TRP-2(R194Q) and TRP-2(H189G), which have lost their DOPAchrome tautomerase activity, failed to modify the HEK cell response to dopamine and hydroquinone. These results suggest that TRP-2 acts on quinone metabolites other than DOPAchrome, e.g., in the catecholamine pathway, and limits their deleterious effects.

MeSH terms

  • Blotting, Western
  • Cell Line, Tumor
  • Cell Survival / drug effects
  • Dopamine / toxicity*
  • Gene Expression Profiling
  • Humans
  • Hydroquinones / toxicity*
  • Intramolecular Oxidoreductases / genetics
  • Intramolecular Oxidoreductases / metabolism*
  • Melanocytes / drug effects*
  • Melanocytes / metabolism
  • Melanocytes / pathology
  • Melanoma, Experimental / metabolism*
  • Mutagenesis, Site-Directed
  • Mutagens / toxicity*

Substances

  • Hydroquinones
  • Mutagens
  • Intramolecular Oxidoreductases
  • dopachrome isomerase
  • Dopamine
  • hydroquinone