| ZHANG, X.W., R.M.K. YU, P.D. JONES, G.K.W. LAM, J.L. NEWSTED, T. GRACIA, M. HECKER, Klára HILSCHEROVÁ, J.T. SANDERSON, R.S.S. WU a J.P. GIESY. Quantitative RT-PCR methods for evaluating toxicant-induced effects on steroidogenesis using the H295R cell line. Environmental Science & Technology. USA: The American Chemical Society, 2005, roč. 39, č. 8, s. 2777-2785. ISSN 0013-936X. |
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@article{827510,
author = {Zhang, X.W. and Yu, R.M.K. and Jones, P.D. and Lam, G.K.W. and Newsted, J.L. and Gracia, T. and Hecker, M. and Hilscherová, Klára and Sanderson, J.T. and Wu, R.S.S. and Giesy, J.P.},
article_location = {USA},
article_number = {8},
keywords = {steroidogenesis; gene expression; in vitro},
language = {eng},
issn = {0013-936X},
journal = {Environmental Science & Technology},
title = {Quantitative RT-PCR methods for evaluating toxicant-induced effects on steroidogenesis using the H295R cell line.},
volume = {39},
year = {2005}
}
TY - JOUR
ID - 827510
AU - Zhang, X.W. - Yu, R.M.K. - Jones, P.D. - Lam, G.K.W. - Newsted, J.L. - Gracia, T. - Hecker, M. - Hilscherová, Klára - Sanderson, J.T. - Wu, R.S.S. - Giesy, J.P.
PY - 2005
TI - Quantitative RT-PCR methods for evaluating toxicant-induced effects on steroidogenesis using the H295R cell line.
JF - Environmental Science & Technology
VL - 39
IS - 8
SP - 2777-2785
EP - 2777-2785
PB - The American Chemical Society
SN - 0013936X
KW - steroidogenesis
KW - gene expression
KW - in vitro
N2 - Gene 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 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 realtime PCR system and quantified based on both a standard curve method using a dilution series of RNA standards and a comparative Ct 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 compoundspecific response profiles, and will provide a sensitive and integrative screen for the effects of chemicals on steroidogenesis.
ER -
ZHANG, X.W., R.M.K. YU, P.D. JONES, G.K.W. LAM, J.L. NEWSTED, T. GRACIA, M. HECKER, Klára HILSCHEROVÁ, J.T. SANDERSON, R.S.S. WU a J.P. GIESY. Quantitative RT-PCR methods for evaluating toxicant-induced effects on steroidogenesis using the H295R cell line. \textit{Environmental Science \&{} Technology}. USA: The American Chemical Society, 2005, roč.~39, č.~8, s.~2777-2785. ISSN~0013-936X.
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