Estimation of the Mechanism of Adrenal Action of Endocrine-Disrupting Compounds Using a Computational Model of Adrenal Steroidogenesis in NCI-H295R Cells

Figure 3

Hierarchical cluster analysis of differential metabolic profiles of 12 steroid hormones by exposure to adrenal toxicity compounds. Adrenal toxicants were classified by using the differential metabolic profiling of 12 steroid hormones. Concentrations of 12 adrenal steroids secreted from NCI-H295R cells were drastically changed by the exposure of adrenal steroidogenesis inhibitors. The 12 adrenal steroid hormones were quantitatively measured by LC-MS/MS simultaneously. Four adrenal vasculotoxic compounds: acrylonitrile: AN, fumaronitrile: FN, salinomycin: SM, and thioguanine: TG, were used as negative control compounds for adrenal steroidogenesis inhibitors. Seven adrenal steroidogenesis inhibitors: aminoglutethimide: AGT, o,p′-DDD: DDD, spironolactone: SP, metyrapone: MP, ketoconazole: KC, miconazole: MC, and daidzein: DZ, showed a characteristic steroid profile each and were classified as each independent singleton at the maximum exposure condition. Exposure concentrations of adrenal toxic compounds were described in brackets of the sample name and the units were prepared as μM. A blue cluster was classified as a group of nonchange samples including negative control compounds and low exposure conditions of adrenal steroidogenesis inhibitors. PREG: pregnenolone, HPREG: 17α-hydroxypregnenolone, DHEA: dehydroepiandrosterone, PROG: progesterone, HPROG: 17α-hydroxyprogesterone, DIONE: androstenedione, DCORTICO: 11-deoxycorticosterone, DCORT: 11-deoxycortisol, CORTICO: corticosterone, CORT: cortisol, ALDO: aldosterone, and TESTO: testosterone.