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

J 2005

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 et. al.

Základní údaje

Originální název

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

Název česky

Kvantitativní RT-PCR metoda pro hodnocení účinku toxikantů na steroidogenezi s využitím buněčné linie H295R

Autoři

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

Vydání

Environmental Science & Technology, USA, The American Chemical Society, 2005, 0013-936X

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

Genetika a molekulární biologie

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Impakt faktor

Impact factor: 4.054

Kód RIV

RIV/00216224:14310/05:00027907

Organizační jednotka

Přírodovědecká fakulta

UT WoS

000228428900053

Klíčová slova česky

steroidogeneze; exprese genů; in vitro

Klíčová slova anglicky

steroidogenesis; gene expression; in vitro

Štítky

GENE EXPRESSION, in vitro, steroidogenesis

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 8. 4. 2009 17:05, doc. Mgr. Klára Hilscherová, Ph.D.

Anotace

ORIG CZ

V originále

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.

Česky

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.

Návaznosti

MSM0021622412, záměr

Název: Interakce mezi chemickými látkami, prostředím a biologickými systémy a jejich důsledky na globální, regionální a lokální úrovni (INCHEMBIOL) (Akronym: INCHEMBIOL)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Interakce mezi chemickými látkami, prostředím a biologickými systémy a jejich důsledky na globální , regionální a lokální úrovni

Citovat

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.

@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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