Type I gamma-GT mRNA is expressed in B16 melanoma and levels correlate with pigmentation

Gamma-glutamyl transpeptidase (gamma-GT), an ectoenzyme involved mainly in glutathione metabolism, is expressed in B16 melanoma cells. B16 melanoma cells under continuous culture conditions show a phenotypic drift from melanotic to amelanotic and re-melanotic stages. We have investigated the regulation of gamma-GT in B16 melanoma cells under such different pigmentary conditions. High levels of gamma-GT messenger RNA (mRNA) and activity were detected in pigmented B16 melanoma cells, whereas in amelanotic B16 melanoma cells the levels were very low. Treatment with lactic acid, a known inhibitor of tyrosinase gene expression, also led to the down-regulation of gamma-GT mRNA and activity. Thus our results indicate that gamma-GT regulation depends on the pigmentation status in pigment cells. We have also assessed the levels of gamma-GT in normal murine melanocytes (melan-a cells). It was seen that melan-a cells express very low levels of gamma-GT. As gamma-GT is known to be regulated in a tissue-specific manner, and is expressed from as many as six promoters giving rise to six different types of mRNAs each having unique 5' ends, we have further investigated the type of gamma-GT mRNA expressed in B16 melanoma and melan-a cells. In this study, we have conclusively demonstrated that type I mRNA transcript of gamma-GT is expressed in B16 melanoma and melan-a cells.