Secretion of cortisol and aldosterone as a vulnerable target for adrenal endocrine disruption — screening of 30 selected chemicals in the human H295R cell model

The adrenal gland is a vulnerable target for toxic insult. Disruption of adrenal steroidogenesis and hormone secretion may cause serious effects on human health. A human in vitro model is needed to predict effects, and elucidate mechanisms of endocrine disruption and adrenal toxicity. The human adrenocortical cell line H295R has been used to screen for effects on sex hormones. Here, we have analyzed the effect of 30 potential endocrine disrupting chemicals on the secretion of cortisol and aldosterone from the H295R cells, using specific ELISA assays. The effect of chemicals was analyzed for basal and forskolin‐ or angiotensin II‐stimulated hormone secretion. The chemicals were tested at the highest concentration where they displayed no evident unspecific cytotoxicity. Quantitative and qualitative differences in effects on hormone secretion were demonstrated for the various chemicals. A subset of the chemicals displayed different effects on cortisol and aldosterone secretion, and in some cases the effects were different between basal and stimulated hormone secretion. Aminoglutethimide, prochloraz, ketoconazole, 6‐hydroxyflavone, imazalil and etomidate had the most marked inhibitory effects on cortisol (with or without forskolin) and ketoconazole, 6‐hydroxyflavone, imazalil and etomidate had the most marked effects on aldosterone (with or without angiotensin II). The results are discussed in terms of known effects, structural similarity and possible mechanisms. We have shown that adrenal steroidogenesis is a vulnerable target for toxic insult and that the H295R assay is a useful in vitro model for screening purposes. Copyright © 2008 John Wiley & Sons, Ltd.

[1]  M. Lam,et al.  Effects of 20 PBDE metabolites on steroidogenesis in the H295R cell line. , 2008, Toxicology letters.

[2]  C. Flück,et al.  Human adrenal corticocarcinoma NCI-H295R cells produce more androgens than NCI-H295A cells and differ in 3beta-hydroxysteroid dehydrogenase type 2 and 17,20 lyase activities. , 2007, The Journal of endocrinology.

[3]  James,et al.  Medical management of Cushing disease. , 2007, Neurosurgical Focus.

[4]  Henner Hollert,et al.  The OECD Validation Program of the H295R Steroidogenesis Assay for the Identification of In Vitro Inhibitors and Inducers of Testosterone and Estradiol Production. Phase 2: Inter-Laboratory Pre-Validation Studies (8 pp) , 2007, Environmental science and pollution research international.

[5]  C. Springall,et al.  Adrenal toxicology: a strategy for assessment of functional toxicity to the adrenal cortex and steroidogenesis , 2007, Journal of applied toxicology : JAT.

[6]  A. Ziółkowska,et al.  Endocrine disruptors and rat adrenocortical function: studies on freshly dispersed and cultured cells. , 2006, International journal of molecular medicine.

[7]  John L Newsted,et al.  Human adrenocarcinoma (H295R) cells for rapid in vitro determination of effects on steroidogenesis: hormone production. , 2006, Toxicology and applied pharmacology.

[8]  E. Ullerås,et al.  Steroidogenic gene expression in H295R cells and the human adrenal gland: adrenotoxic effects of lindane in vitro , 2006, Journal of applied toxicology : JAT.

[9]  J. Hinson,et al.  Effects of endocrine-disrupting chemicals on adrenal function. , 2006, Best practice & research. Clinical endocrinology & metabolism.

[10]  Rolf W. Hartmann,et al.  The adrenocortical tumor cell line NCI-H295R as an in vitro screening system for the evaluation of CYP11B2 (aldosterone synthase) and CYP11B1 (steroid-11β-hydroxylase) inhibitors , 2005, The Journal of Steroid Biochemistry and Molecular Biology.

[11]  Lih-Ann Li,et al.  PCB126 induces differential changes in androgen, cortisol, and aldosterone biosynthesis in human adrenocortical H295R cells. , 2005, Toxicological sciences : an official journal of the Society of Toxicology.

[12]  B. Schimmer,et al.  Adrenocortical cell lines , 2004, Molecular and Cellular Endocrinology.

[13]  P. Harvey,et al.  The adrenal cortex and steroidogenesis as cellular and molecular targets for toxicity: critical omissions from regulatory endocrine disrupter screening strategies for human health? , 2003, Journal of applied toxicology : JAT.

[14]  Martin van den Berg,et al.  Induction and inhibition of aromatase (CYP19) activity by various classes of pesticides in H295R human adrenocortical carcinoma cells. , 2002, Toxicology and applied pharmacology.

[15]  Johansson Mk,et al.  Effects of 3-MeSO2-DDE and some CYP inhibitors on glucocorticoid steroidogenesis in the H295R human adrenocortical carcinoma cell line. , 2002 .

[16]  J. Goldman,et al.  Alterations in ovarian follicular progesterone secretion by elevated exposures to the drinking water disinfection by-product dibromoacetic acid: examination of the potential site(s) of impact along the steroidogenic pathway. , 2002, Toxicology.

[17]  T. Rosol,et al.  Adrenal Gland: Structure, Function, and Mechanisms of Toxicity , 2001, Toxicologic pathology.

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

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

[20]  J. Mathis,et al.  Human NCI-H295 adrenocortical carcinoma cells: a model for angiotensin-II-responsive aldosterone secretion. , 1993, Endocrinology.

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

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

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

[24]  W. E. Ribelin.  The effects of drugs and chemicals upon the structure of the adrenal gland. , 1984, Fundamental and applied toxicology : official journal of the Society of Toxicology.

[25]  I. Ledingham,et al.  INFLUENCE OF SEDATION ON MORTALITY IN CRITICALLY ILL MULTIPLE TRAUMA PATIENTS , 1983, The Lancet.

[26]  C. Mélot,et al.  SYSTEMIC AND PULMONARY HAEMODYNAMIC EFFECTS OF SOMATOSTATIN , 1981, The Lancet.

[27]  J. Giesy,et al.  Novel trends in endocrine disruptor testing: the H295R Steroidogenesis Assay for identification of inducers and inhibitors of hormone production , 2008, Analytical and bioanalytical chemistry.

[28]  J. T. Sanderson,et al.  Effects of 3-MeSO2-DDE and some CYP inhibitors on glucocorticoid steroidogenesis in the H295R human adrenocortical carcinoma cell line. , 2002, Toxicology in vitro : an international journal published in association with BIBRA.

[29]  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.