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Article: Quantitative RT-PCR methods for evaluating toxicant-induced effects on steroidogenesis using the H295R cell line
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TitleQuantitative RT-PCR methods for evaluating toxicant-induced effects on steroidogenesis using the H295R cell line
 
AuthorsZhang, X1 3
Yu, RMK3
Jones, PD1
Lam, GKW3
Newsted, JL2
Gracia, T1
Hecker, M1
Hilscherova, K1
Sanderson, JT4
Wu, RSS3
Giesy, JP1 3
 
KeywordsMolecular Sequence Numbers
 
Issue Date2005
 
PublisherAmerican Chemical Society. The Journal's web site is located at http://pubs.acs.org/est
 
CitationEnvironmental Science And Technology, 2005, v. 39 n. 8, p. 2777-2785 [How to Cite?]
DOI: http://dx.doi.org/10.1021/es048679k
 
AbstractGene expression profiles show considerable promise for the evaluation of the toxic potential of environmental contaminants. For example, any alterations in the pathways of steroid synthesis or breakdown have the potential to cause endocrine disruption. Therefore monitoring these pathways can provide information relative to a chemical's ability to impact endocrine function. One approach to monitoring these pathways has been to use a human adrenocortical carcinoma cell line (H295R) that expresses all the key enzymes necessary for steroidogenesis. In this study we have further developed these methods using accurate and specific quantification methods utilizing molecular beacon-based quantitative RT-PCR (Q-RT-PCR). The assay system was used to analyze the expression patterns of 11 steroidogenic genes in H295R cells. The expression of gene transcripts was measured using a real-time PCR system and quantified based on both a standard curve method using a dilution series of RNA standards and a comparative C t method. To validate the optimized method, cells were exposed to specific and nonspecific model compounds (inducers and inhibitors of various steroidogenic enzymes) for gene expression profiling. Similar gene expression profiles were exhibited by cells treated with chemicals acting through common mechanisms of action. Overall, our findings demonstrated that the present assay can facilitate the development of compound-specific response profiles, and will provide a sensitive and integrative screen for the effects of chemicals on steroidogenesis. © 2005 American Chemical Society.
 
ISSN0013-936X
2013 Impact Factor: 5.481
 
DOIhttp://dx.doi.org/10.1021/es048679k
 
ISI Accession Number IDWOS:000228428900053
 
ReferencesReferences in Scopus
 
DC FieldValue
dc.contributor.authorZhang, X
 
dc.contributor.authorYu, RMK
 
dc.contributor.authorJones, PD
 
dc.contributor.authorLam, GKW
 
dc.contributor.authorNewsted, JL
 
dc.contributor.authorGracia, T
 
dc.contributor.authorHecker, M
 
dc.contributor.authorHilscherova, K
 
dc.contributor.authorSanderson, JT
 
dc.contributor.authorWu, RSS
 
dc.contributor.authorGiesy, JP
 
dc.date.accessioned2010-09-17T10:57:12Z
 
dc.date.available2010-09-17T10:57:12Z
 
dc.date.issued2005
 
dc.description.abstractGene expression profiles show considerable promise for the evaluation of the toxic potential of environmental contaminants. For example, any alterations in the pathways of steroid synthesis or breakdown have the potential to cause endocrine disruption. Therefore monitoring these pathways can provide information relative to a chemical's ability to impact endocrine function. One approach to monitoring these pathways has been to use a human adrenocortical carcinoma cell line (H295R) that expresses all the key enzymes necessary for steroidogenesis. In this study we have further developed these methods using accurate and specific quantification methods utilizing molecular beacon-based quantitative RT-PCR (Q-RT-PCR). The assay system was used to analyze the expression patterns of 11 steroidogenic genes in H295R cells. The expression of gene transcripts was measured using a real-time PCR system and quantified based on both a standard curve method using a dilution series of RNA standards and a comparative C t method. To validate the optimized method, cells were exposed to specific and nonspecific model compounds (inducers and inhibitors of various steroidogenic enzymes) for gene expression profiling. Similar gene expression profiles were exhibited by cells treated with chemicals acting through common mechanisms of action. Overall, our findings demonstrated that the present assay can facilitate the development of compound-specific response profiles, and will provide a sensitive and integrative screen for the effects of chemicals on steroidogenesis. © 2005 American Chemical Society.
 
dc.description.natureLink_to_subscribed_fulltext
 
dc.identifier.citationEnvironmental Science And Technology, 2005, v. 39 n. 8, p. 2777-2785 [How to Cite?]
DOI: http://dx.doi.org/10.1021/es048679k
 
dc.identifier.doihttp://dx.doi.org/10.1021/es048679k
 
dc.identifier.epage2785
 
dc.identifier.isiWOS:000228428900053
 
dc.identifier.issn0013-936X
2013 Impact Factor: 5.481
 
dc.identifier.issue8
 
dc.identifier.pmid15884376
 
dc.identifier.scopuseid_2-s2.0-20244384266
 
dc.identifier.spage2777
 
dc.identifier.urihttp://hdl.handle.net/10722/92789
 
dc.identifier.volume39
 
dc.languageeng
 
dc.publisherAmerican Chemical Society. The Journal's web site is located at http://pubs.acs.org/est
 
dc.publisher.placeUnited States
 
dc.relation.ispartofEnvironmental Science and Technology
 
dc.relation.referencesReferences in Scopus
 
dc.subjectMolecular Sequence Numbers
 
dc.titleQuantitative RT-PCR methods for evaluating toxicant-induced effects on steroidogenesis using the H295R cell line
 
dc.typeArticle
 
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<contributor.author>Newsted, JL</contributor.author>
<contributor.author>Gracia, T</contributor.author>
<contributor.author>Hecker, M</contributor.author>
<contributor.author>Hilscherova, K</contributor.author>
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Author Affiliations
  1. Michigan State University
  2. ENTRIX Inc.
  3. City University of Hong Kong
  4. Utrecht University