Quantitative RT-PCR methods for evaluating toxicant-induced effects on steroidogenesis using the H295R cell line.

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 and 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, vol. 39, No 8, p. 2777-2785. ISSN 0013-936X.

Basic information

• Original name: Quantitative RT-PCR methods for evaluating toxicant-induced effects on steroidogenesis using the H295R cell line.
• Name in Czech: Kvantitativní RT-PCR metoda pro hodnocení účinku toxikantů na steroidogenezi s využitím buněčné linie H295R
• Authors: ZHANG, X.W. (156 China), R.M.K. YU (156 China), P.D. JONES (840 United States of America), G.K.W. LAM (156 China), J.L. NEWSTED (840 United States of America), T. GRACIA (170 Colombia), M. HECKER (276 Germany), Klára HILSCHEROVÁ (203 Czech Republic, guarantor), J.T. SANDERSON (528 Netherlands), R.S.S. WU (156 China) and J.P. GIESY (840 United States of America).
• Edition: Environmental Science & Technology, USA, The American Chemical Society, 2005, 0013-936X.

Other information

• Original language: English
• Type of outcome: Article in a journal
• Field of Study: Genetics and molecular biology
• Country of publisher: United States of America
• Confidentiality degree: is not subject to a state or trade secret
• Impact factor: Impact factor: 4.054
• RIV identification code: RIV/00216224:14310/05:00027907
• Organization unit: Faculty of Science
• UT WoS: 000228428900053
• Keywords (in Czech): steroidogeneze; exprese genů; in vitro
• Keywords in English: steroidogenesis; gene expression; in vitro
• Tags: GENE EXPRESSION, in vitro, steroidogenesis
• Tags: International impact, Reviewed

Abstract

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.

Abstract (in Czech)

V této práci je shrnuto ovlivnění profilu odpovědí 11 steroidogenních enzymů na modelové induktory a inhibitory steroidogeneze v in vitro modelovém systému pomocí real-time PCR.

Links

• MSM0021622412, plan (intention): Name: Interakce mezi chemickými látkami, prostředím a biologickými systémy a jejich důsledky na globální, regionální a lokální úrovni (INCHEMBIOL) (Acronym: INCHEMBIOL)

Investor: Ministry of Education, Youth and Sports of the CR, Interactions among the chemicals, environment and biological systems and their consequences on the global, regional and local scales (INCHEMBIOL)