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Article: Human adrenocarcinoma (H295R) cells for rapid in vitro determination of effects on steroidogenesis: Hormone production
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TitleHuman adrenocarcinoma (H295R) cells for rapid in vitro determination of effects on steroidogenesis: Hormone production
 
AuthorsHecker, M4
Newsted, JL2
Murphy, MB4 3
Higley, EB4
Jones, PD4
Wu, R3
Giesy, JP4 3 1
 
KeywordsChemical screening
Endocrine disruption
Steroid hormones
Steroidogenesis
 
Issue Date2006
 
PublisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/locate/taap
 
CitationToxicology And Applied Pharmacology, 2006, v. 217 n. 1, p. 114-124 [How to Cite?]
DOI: http://dx.doi.org/10.1016/j.taap.2006.07.007
 
AbstractTo identify and prioritize chemicals that may alter steroidogenesis, an in vitro screening assay based on measuring alterations in hormone production was developed using the H295R human adrenocortical carcinoma cell line. Previous studies indicated that this cell line was useful to screen for effects on gene expression of steroidogenic enzymes. This study extended that work to measure the integrated response on production of testosterone (T), estradiol (E2), and progesterone/pregnenolone (P) using an ELISA. Under optimized culture and experimental conditions, the basal release of P, T and E2 into the medium was 7.0 ± 1.2 ng/ml, 1.6 ± 0.4 ng/ml, and 0.51 ± 0.13 ng/ml, respectively. Model chemicals with different modes of action on steroidogenic systems were tested. Exposure to forskolin resulted in dose-dependent increases in all three hormones with the greatest relative increase being observed for E2. This differed from cells exposed to prochloraz or ketoconazole where P concentrations increased while T and E2 concentrations decreased in a dose-dependent manner. In cells exposed to fadrozole, E2 decreased in a dose-dependent manner while T and P only decreased at the greatest dose tested. Aminoglutethimide decreased P and E2 concentrations but increased T concentrations. Vinclozolin reduced both P and T but resulted in a slight increase in E2. The alteration in the patterns of hormone production in the H295R assay was consistent with the modes of action of the chemicals and was also consistent with observed effects of these chemicals in animal models. Based on these results, the H295R in vitro system has potential for high throughput screening to not only characterize the effects of chemicals on endocrine systems but also to prioritize chemicals for additional testing. © 2006 Elsevier Inc. All rights reserved.
 
ISSN0041-008X
2013 Impact Factor: 3.630
 
DOIhttp://dx.doi.org/10.1016/j.taap.2006.07.007
 
ISI Accession Number IDWOS:000241937100013
 
ReferencesReferences in Scopus
 
DC FieldValue
dc.contributor.authorHecker, M
 
dc.contributor.authorNewsted, JL
 
dc.contributor.authorMurphy, MB
 
dc.contributor.authorHigley, EB
 
dc.contributor.authorJones, PD
 
dc.contributor.authorWu, R
 
dc.contributor.authorGiesy, JP
 
dc.date.accessioned2010-09-17T10:56:53Z
 
dc.date.available2010-09-17T10:56:53Z
 
dc.date.issued2006
 
dc.description.abstractTo identify and prioritize chemicals that may alter steroidogenesis, an in vitro screening assay based on measuring alterations in hormone production was developed using the H295R human adrenocortical carcinoma cell line. Previous studies indicated that this cell line was useful to screen for effects on gene expression of steroidogenic enzymes. This study extended that work to measure the integrated response on production of testosterone (T), estradiol (E2), and progesterone/pregnenolone (P) using an ELISA. Under optimized culture and experimental conditions, the basal release of P, T and E2 into the medium was 7.0 ± 1.2 ng/ml, 1.6 ± 0.4 ng/ml, and 0.51 ± 0.13 ng/ml, respectively. Model chemicals with different modes of action on steroidogenic systems were tested. Exposure to forskolin resulted in dose-dependent increases in all three hormones with the greatest relative increase being observed for E2. This differed from cells exposed to prochloraz or ketoconazole where P concentrations increased while T and E2 concentrations decreased in a dose-dependent manner. In cells exposed to fadrozole, E2 decreased in a dose-dependent manner while T and P only decreased at the greatest dose tested. Aminoglutethimide decreased P and E2 concentrations but increased T concentrations. Vinclozolin reduced both P and T but resulted in a slight increase in E2. The alteration in the patterns of hormone production in the H295R assay was consistent with the modes of action of the chemicals and was also consistent with observed effects of these chemicals in animal models. Based on these results, the H295R in vitro system has potential for high throughput screening to not only characterize the effects of chemicals on endocrine systems but also to prioritize chemicals for additional testing. © 2006 Elsevier Inc. All rights reserved.
 
dc.description.naturelink_to_subscribed_fulltext
 
dc.identifier.citationToxicology And Applied Pharmacology, 2006, v. 217 n. 1, p. 114-124 [How to Cite?]
DOI: http://dx.doi.org/10.1016/j.taap.2006.07.007
 
dc.identifier.doihttp://dx.doi.org/10.1016/j.taap.2006.07.007
 
dc.identifier.eissn1096-0333
 
dc.identifier.epage124
 
dc.identifier.isiWOS:000241937100013
 
dc.identifier.issn0041-008X
2013 Impact Factor: 3.630
 
dc.identifier.issue1
 
dc.identifier.pmid16962624
 
dc.identifier.scopuseid_2-s2.0-33750351770
 
dc.identifier.spage114
 
dc.identifier.urihttp://hdl.handle.net/10722/92778
 
dc.identifier.volume217
 
dc.languageeng
 
dc.publisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/locate/taap
 
dc.publisher.placeUnited States
 
dc.relation.ispartofToxicology and Applied Pharmacology
 
dc.relation.referencesReferences in Scopus
 
dc.subjectChemical screening
 
dc.subjectEndocrine disruption
 
dc.subjectSteroid hormones
 
dc.subjectSteroidogenesis
 
dc.titleHuman adrenocarcinoma (H295R) cells for rapid in vitro determination of effects on steroidogenesis: Hormone production
 
dc.typeArticle
 
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<description.abstract>To identify and prioritize chemicals that may alter steroidogenesis, an in vitro screening assay based on measuring alterations in hormone production was developed using the H295R human adrenocortical carcinoma cell line. Previous studies indicated that this cell line was useful to screen for effects on gene expression of steroidogenic enzymes. This study extended that work to measure the integrated response on production of testosterone (T), estradiol (E2), and progesterone/pregnenolone (P) using an ELISA. Under optimized culture and experimental conditions, the basal release of P, T and E2 into the medium was 7.0 &#177; 1.2&#160;ng/ml, 1.6 &#177; 0.4&#160;ng/ml, and 0.51 &#177; 0.13&#160;ng/ml, respectively. Model chemicals with different modes of action on steroidogenic systems were tested. Exposure to forskolin resulted in dose-dependent increases in all three hormones with the greatest relative increase being observed for E2. This differed from cells exposed to prochloraz or ketoconazole where P concentrations increased while T and E2 concentrations decreased in a dose-dependent manner. In cells exposed to fadrozole, E2 decreased in a dose-dependent manner while T and P only decreased at the greatest dose tested. Aminoglutethimide decreased P and E2 concentrations but increased T concentrations. Vinclozolin reduced both P and T but resulted in a slight increase in E2. The alteration in the patterns of hormone production in the H295R assay was consistent with the modes of action of the chemicals and was also consistent with observed effects of these chemicals in animal models. Based on these results, the H295R in vitro system has potential for high throughput screening to not only characterize the effects of chemicals on endocrine systems but also to prioritize chemicals for additional testing. &#169; 2006 Elsevier Inc. All rights reserved.</description.abstract>
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Author Affiliations
  1. University of Saskatchewan
  2. ENTRIX Inc.
  3. City University of Hong Kong
  4. National Food Safety and Toxicology Center