Two Panels of Steroid Receptor Luciferase Reporter Cell Lines for Compound Profiling

• DOI: 10.2174/138620711795222446
• Publication Date: 2011-11-01T15:25:38Z
• Authors: David Sedlak; Aileen Paguio; Petr Bartunek
• Keywords: Steroid hormone receptor; Hormone response element

Steroid hormone receptors represent a major target in drug discovery. As ligand inducible transcription factors, their activity can be modulated by small lipophilic molecules. Here we describe two panels of potent and selective luciferase reporter cell lines based on cells with low endogenous steroid receptor (U2OS). The contain for estrogen α β, androgen, glucocorticoid, mineralocorticoid, progesterone receptors. In the first panel, activation either synthetic, response elements containing promoter or viral is mediated full-length second panel expression chimeric receptor, which was created replacement N-terminal part molecule Gal4 DBD that binds to multiple UAS sites promoter. Both were extensively characterized profiling 28 ligands dose manner agonist antagonist mode. We have analyzed compared responses tested from both concluded general systems generated similar qualitative terms potency, efficacy, partial agonism/antagonism, mixed agonistic/antagonistic profiles rank potencies well conserved between panels. However, also identified some artifacts introduced Gal4/LBD assays contrast counterparts. Keeping mind advantages drawbacks each format, these powerful tools large compound libraries (HTS) detailed study mechanisms compounds exert biological effects. Keywords: U2OS, cell-based assay, binding domain, Gal4, nuclear HTS, discovery, Cell Lines, posttranslational modifications, DNA vectors