Effects of 3-MeSO2-DDE and some CYP inhibitors on glucocorticoid steroidogenesis in the H295R human adrenocortical carcinoma cell line.

Abstract The formation of steroids in the H295R human adrenocortical carcinoma cell line was analysed by HPLC or RIA, and based on these data the apparent catalytic activities of CYP11A, CYP17, CYP21 and CYP11B1 in this cell line were calculated. The environmental pollutant 3-methylsulfonyl-DDE (3-MeSO2-DDE) and the cytochrome P450 (CYP) inhibitors ketoconazole, metyrapone and aminoglutethimide were studied for their effects on the steroid formation. Metyrapone (IC50 of 1 μ m ) and 3-MeSO2-DDE (10 μ m : 66±10% of control) were found to inhibit the apparent CYP11B1 activity. Ketoconazole inhibited all enzymes examined with the greatest effects on CYP11B1 (IC50 of 2.5 μ m ). Aminoglutethimide was examined only for effects on CYP11A activity and was shown to inhibit pregnenolone formation (20 μ m : 61±4% of control). The possibility of studying all CYP enzymes in the corticosteroidogenesis makes this cell line a valuable test system to examine effects of chemicals, such as suspected endocrine disruptors, on the human glucocorticoid hormone synthesis. The inhibition of cortisol formation by 3-MeSO2-DDE supports an interaction with the active site of CYP11B1, as previously reported in mouse adrenocortical Y1 cells. In mice, this interaction led to metabolic activation and a high adrenotoxicity of 3-MeSO2-DDE. Therefore studies on the adrenotoxicity of 3-MeSO2-DDE in humans are needed.

[1]  T. Triche,et al.  Establishment and characterization of a human adrenocortical carcinoma cell line that expresses multiple pathways of steroid biosynthesis. , 1990, Cancer research.

[2]  B. Lund,et al.  In vitro bioactivation of the environmental pollutant 3-methylsulphonyl-2, 2-bis(4-chlorophenyl)-1,1-dichloroethene in the human adrenal gland. , 1994, Toxicology letters.

[3]  D. Feldman,et al.  Ketoconazole and other imidazole derivatives as inhibitors of steroidogenesis. , 1986, Endocrine reviews.

[4]  J. Dunn,et al.  The effect of ketoconazole on steroidogenesis. II. Adrenocortical enzyme activity in vitro. , 1988, Research communications in chemical pathology and pharmacology.

[5]  W. Beierwaltes,et al.  Adrenal cortical 11 beta-hydroxylase and side-chain cleavage enzymes. Requirement for the A- or B-pyridyl ring in metyrapone for inhibition. , 1985, Journal of steroid biochemistry.

[6]  M. Isamu,et al.  Effect of ketoconazole (an imidazole antimycotic agent) and other inhibitors of steroidogenesis on cytochrome P450-catalyzed reactions , 1986 .

[7]  D. Feldman,et al.  Ketoconazole blocks adrenal steroidogenesis by inhibiting cytochrome P450-dependent enzymes. , 1983, The Journal of clinical investigation.

[8]  R. Fraser,et al.  Inhibition of corticosteroid 11 beta hydroxylation in bovine zona fasciculata cells by the potassium entry blocker 4-aminopyridine. , 1986, Journal of steroid biochemistry.

[9]  S. Szabó,et al.  Adrenal Gland: Chemically Induced Structural and Functional Changes in the Cortex , 1989, Toxicologic pathology.

[10]  A. Bergman,et al.  Metabolic activation and toxicity of a DDT-metabolite, 3-methylsulphonyl-DDE, in the adrenal zona fasciculata in mice. , 1988, Chemico-biological interactions.

[11]  H. Akaza,et al.  YM116, 2-(1H-imidazol-4-ylmethyl)-9H-carbazole, decreases adrenal androgen synthesis by inhibiting C17-20 lyase activity in NCI-H295 human adrenocortical carcinoma cells. , 1999, Japanese journal of pharmacology.

[12]  J. Lund,et al.  Novel Involvement of a Mitochondrial Steroid Hydroxylase (P450c11) in Xenobiotic Metabolism (*) , 1995, The Journal of Biological Chemistry.

[13]  F. Beuschlein,et al.  Steroidogenic acute regulatory protein mRNA expression in adrenal tumours. , 2000, European journal of endocrinology.

[14]  O. H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.

[15]  F. Beuschlein,et al.  Aminoglutethimide suppresses adrenocorticotropin receptor expression in the NCI-h295 adrenocortical tumor cell line. , 1998, The Journal of endocrinology.

[16]  D. Feldman,et al.  Inhibition of steroidogenic cytochrome P-450 enzymes in rat testis by ketoconazole and related imidazole anti-fungal drugs. , 1985, Journal of steroid biochemistry.

[17]  M. Kawamura,et al.  Procaine inhibits cyclic AMP-induced steroidogenesis in isolated bovine adrenocortical cells. , 1990, Japanese journal of pharmacology.

[18]  A. Bergman,et al.  Adrenocortical toxicity of 3-methylsulphonyl-DDE; 3: Studies in fetal and suckling mice. , 1992, Reproductive toxicology.

[19]  M. Levell,et al.  Inhibition of human adrenal steroidogenic enzymes in vitro by imidazole drugs including ketoconazole. , 1989, Journal of steroid biochemistry.

[20]  H. Rodríguez-Martínez,et al.  Adrenocortical toxicity of 3-methylsulfonyl-DDE in mice. II. Mitochondrial changes following ecologically relevant doses. , 1991, Fundamental and applied toxicology : official journal of the Society of Toxicology.

[21]  H. Bruining,et al.  Differential effects of the imidazole derivatives etomidate, ketoconazole and miconazole and of metyrapone on the secretion of cortisol and its precursors by human adrenocortical cells. , 1987, The Journal of pharmacology and experimental therapeutics.

[22]  A. Bergman,et al.  Synthesis of methanesulfonyl derivatives of 2,2-bis (4-chlorophenyl)-1,1-dichloroethylene (p,p'-DDE), present in seal from the Baltic. , 1977, Acta chemica Scandinavica. Series B: Organic chemistry and biochemistry.

[23]  K. Norén,et al.  Methylsulfonyl metabolites of PCBs and DDE in human milk in Sweden, 1972-1992. , 1996, Environmental health perspectives.

[24]  A. Bergman,et al.  Structure-activity relationship for inhibition of CYP11B1-dependent glucocorticoid synthesis in Y1 cells by aryl methyl sulfones. , 1998, Pharmacology & toxicology.

[25]  D. Hum,et al.  Regulation of steroidogenesis in NCI-H295 cells: a cellular model of the human fetal adrenal. , 1993, Molecular endocrinology.

[26]  D. Smith,et al.  Worldwide trends in DDT levels in human breast milk. , 1999, International journal of epidemiology.

[27]  G. Gillies,et al.  Stress, stress hormones and the immune system , 1997 .

[28]  I. Bird,et al.  The NCI-H295 cell line: a pluripotent model for human adrenocortical studies , 1994, Molecular and Cellular Endocrinology.

[29]  P. White,et al.  Inhibition of adrenal steroidogenesis by the anesthetic etomidate. , 1984, The New England journal of medicine.

[30]  K. Norén,et al.  Methylsulfonyl metabolites of PCBs and DDE in human tissues. , 1997, Environmental health perspectives.

[31]  D. Greenblatt,et al.  Effects of the Antifungal Agents on Oxidative Drug Metabolism , 2000, Clinical pharmacokinetics.

[32]  L. Fishman,et al.  Biosynthesis and metabolism of steroid hormones by human adrenal carcinomas. , 2000, Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas.

[33]  H. Colby.  In vitro assessment of adrenocortical toxicity. , 1994, Journal of pharmacological and toxicological methods.

[34]  L. Fishman,et al.  Dual sites of inhibition by metyrapone of human adrenal steroidogenesis: correlation of in vivo and in vitro studies. , 1976, The Journal of clinical endocrinology and metabolism.

[35]  J. Castro,et al.  Benznidazole-induced ultrastructural alterations in rat adrenal cortex. Mechanistic studies. , 1992, Toxicology.

[36]  N. Hanley,et al.  Regulation of human adrenal carcinoma cell (NCI-H295) production of C19 steroids. , 1993, The Journal of clinical endocrinology and metabolism.

[37]  B. Brunström,et al.  Developmental and reproductive toxicity of persistent environmental pollutants. , 1998, Archives of toxicology. Supplement. = Archiv fur Toxikologie. Supplement.

[38]  P. Bournot,et al.  18-Hydroxylation in the Y-1 adrenal cell line: Response to acth and to culture conditions , 1992, The Journal of Steroid Biochemistry and Molecular Biology.