Test No. 458: Stably Transfected Human Androgen Receptor Transcriptional Activation Assay for Detection of Androgenic Agonist and Antagonist Activity of Chemicals

This Test Guideline describes an in vitro assay providing the methodology of Stably Transfected Transactivation to detect Androgen Receptor Agonists and Antagonists (AR STTA assays). The TA assay using a reporter gene technique is an in vitro tool that provides mechanistic data. The assay is used to establish signal activation or blocking of the androgen receptor caused by a ligand. Some chemicals may, in a cell type-dependent manner, display both agonist and antagonist activity and are known as selective androgen receptor modulators. Following the ligand binding, the receptor-ligand complex translocates to the nucleus where it binds specific DNA response elements and transactivates a firefly luciferase reporter gene, resulting in an increased cellular expression of the luciferase enzyme. Luciferin is a substrate that is transformed by the luciferase enzyme to a bioluminescence product that can be quantitatively measured with a luminometer. Luciferase activity can be evaluated quickly and inexpensively with a number of commercially available test kits. The test system provided in this Test Guideline utilises the AR-EcoScreenTM cell line.

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