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PROFILING GENE EXPRESSION IN HUMAN H295R ADRENOCORTICAL CARCINOMA CELLS AND RAT TESTES TO IDENTIFY PATHWAYS OF TOXICITY FOR CONAZOLE FUNGICIDES
Citation:
Ren, H, J E. Schmid, J. Retief, Y. Turpaz, X. Zhang, P. Jones, J. L. Newsted, J P. Giesy, D C. Wolf, C R. Wood, W Bao, AND D J. Dix. PROFILING GENE EXPRESSION IN HUMAN H295R ADRENOCORTICAL CARCINOMA CELLS AND RAT TESTES TO IDENTIFY PATHWAYS OF TOXICITY FOR CONAZOLE FUNGICIDES. Presented at Society of Toxicology, New Orleans, LA, March 6-10, 2005.
Description:
Profiling Gene Expression in Human H295R Adrenocortical Carcinoma Cells and Rat Testes to Identify Pathways of Toxicity for Conazole Fungicides
Ren1, H., Schmid1, J., Retief2, J., Turpaz2, Y.,Zhang3, X.,Jones3, P., Newsted3, J.,Giesy3, J., Wolf1, D.,Wood1, C., Bao1, W., Dix1, D.
1US Environmental Protection Agency, RTP, NC 27711; 2Affymetrix Inc., Santa Clara, CA 95051;3Michigan State University, East Lansing, MI 48824
Conazoles are widely used agricultural and pharmaceutical fungicides which inhibit sterol and steroid biosynthesis. Studies with 3 agricultural conazoleswere conducted by in vitro exposure of H295Rcells and in vivo exposure of Wistar-Han rats. H295R cells express key steroidogenic enzymes and represent an alternativeto the rat testis for assessing endocrine disruption. We used quantitative PCR (qPCR) and DNA microarraysto profile gene expression and identify common effects and toxicity pathways. H295R cells were cultured with 1, 3, 10, 30 or 100 ?M of each conazole for 48-72 hours. Adult rats were fed myclobutanil (100, 500, 2000 ppm), triadimefon(100, 500, 1800ppm), propiconazole (100, 500, 2500 ppm), or vehicle-added rodent chowforfour days. Expression of 9 steroidogenic genes was measured by qPCR for all H295R dose groups; Affymetrix Human U133plus2.0GeneChips were used for the 3 higher doses of each conazole.Rat gene expression for all dose groups was profiled using Affymetrix Rat2302.0 GeneChips. Resulting data were log-transformed and analyzed by multi-way ANOVA to determine interactions between the chemical and the in vitro and in vivo systems. H295R qPCR results indicated that gene expression of Cyp17, 3?HSD2, 17?HSD1 and Cyp19 are affected by conazole treatment. Microarray results also show similar effects on these steroidogenic genes and indicated the same trend in orthologous rat genes. Broader analysis of gene expression changes in H295R cells and rat testisidentified common biological pathways, including programmed cell death, cell cycle, lipid metabolism and steroid metabolism. Further analysis of results from both H295R cells and rat testis is underway to reveal similar effects after conazole exposure and potential common modes of toxic action. [This abstract does not necessarily reflect EPA policy]