Human epidermal cell protein responses to arsenite treatment in culture

Abstract

Study of the responses of target cells in culture is anticipated to help understand the mechanisms by which inorganic arsenic causes pathological effects in vivo. Treatment of human epidermal cells with arsenic has been shown to produce a myriad of changes in gene transcription. Present work focused on finding the extent of arsenite-induced changes in the protein pattern and whether global effects on protein sulfhydryls were evident. First, examining the profile of protein expression by two-dimensional gel electrophoresis indicated that approximate to 40% of the 300 distinct protein spots that were monitored changed by at least two-fold in amount all through a 9-day exposure period. Second, examining soluble extracts of the treated cells by Activated Thiol Sepharose column chromatography gave little indication of change in the overall protein thiol content. Finally, among the 10 proteins identified that showed prominent changes in amount as a result of treatment for I or 4 days, enzymes of the glycolytic pathway were seen to be substantially elevated as a result of treatment, suggesting decreased utilization by the cells of oxidative phosphorylation. Since these changes were more conspicuous at the protein level than in previous transcriptional studies, the results emphasize the importance of proteomic analysis to complement transcriptional analysis of cell responses to perturbation by arsenic. (c) 2005 Elsevier Ireland Ltd. All rights reserved.