Optimization and prevalidation of the in vitro ERα CALUX method to test estrogenic and antiestrogenic activity of compounds

被引：66

作者：

van der Burg, Bart ^{[1
]}

Winter, Roos ^{[1
]}

Weimer, Marc ^{[2
]}

Berckmans, Pascale ^{[3
]}

Suzuki, Go ^{[4
]}

Gijsbers, Linda ^{[5
]}

Jonas, Arjen ^{[1
]}

van der Linden, Sander ^{[1
]}

Witters, Hilda ^{[3
]}

Aarts, Jac ^{[5
]}

Legler, Juliette ^{[6
]}

Kopp-Schneider, Annette ^{[2
]}

Bremer, Susanne ^{[7
]}

机构：

[1] BioDetect Syst BV, NL-1098 XH Amsterdam, Netherlands

[2] German Canc Res Ctr, Dept Biostat, D-6900 Heidelberg, Germany

[3] Flemish Inst Technol Res, Mol, Belgium

[4] Ehime Univ, Ctr Marine Environm Studies, Ehima, Japan

[5] Wageningen Univ, Div Toxicol, Wageningen, Netherlands

[6] Vrije Univ Amsterdam, Inst Environm Studies, Amsterdam, Netherlands

[7] European Ctr Validat Alternat Methods, Ispra, Italy

来源：

REPRODUCTIVE TOXICOLOGY | 2010年 / 30卷 / 01期

关键词：

Estrogen; Antiestrogen; Reporter gene assay; Uterotrophic assay; In vitro; Validation; CELL-LINE; TRANSACTIVATION ASSAY; ENDOCRINE DISRUPTORS; BIOASSAYS; ANDROGEN; CHEMICALS; PANEL; BETA;

D O I：

10.1016/j.reprotox.2010.04.007

中图分类号：

Q [生物科学];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Estrogenicity of chemicals has received significant attention and is linked to endocrine-disrupting activities. However, there is a paucity of validated methods to assess estrogenicity in vitro. We have established a robust method to test estrogenic and antiestrogenic activity of compounds in vitro, as an alternative to using animal models such as the uterotrophic assay. To this end we optimized protocols to be used in combination with CALUX reporter gene assays and carried out an in house prevalidation, followed by two rounds of tests to establish transferability. Problems in the initial test with transferability were solved by isolation of a novel cell clone of the ER alpha CALUX line with greatly improved stability and luciferase levels. This cell line proved to be a very suitable and reliable predictor of estrogenicity of chemicals and was able to readily rank a range of chemicals on the basis of their EC50 values. (C) 2010 Elsevier Inc. All rights reserved.

引用

页码：73 / 80

页数：8

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