Abstract
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 activity (U2OS). The panels contain reporter cell lines for estrogen receptors α and β, androgen, glucocorticoid, mineralocorticoid, and progesterone receptors. In the first panel, the activation of either synthetic, steroid response elements containing promoter or viral promoter is mediated by full-length steroid receptors. The second panel is based on the expression of the chimeric receptor, which was created by the replacement of the N-terminal part of the molecule by Gal4 DBD and that binds to multiple UAS sites in the reporter promoter. Both panels were extensively characterized by profiling 28 ligands in dose response manner in agonist and antagonist mode. We have analyzed and compared the responses to tested ligands from both panels and concluded that in general both systems generated similar qualitative response in terms of potency, efficacy, partial agonism/antagonism, mixed agonistic/antagonistic profiles and the rank of potencies was well conserved between both panels. However, we have also identified some artifacts introduced by the Gal4/LBD reporter assays in contrast to their full-length receptor reporter counterparts. Keeping in mind the advantages and drawbacks of each reporter format, these cell lines represent powerful and selective tools for profiling large compound libraries (HTS) and for detailed study of mechanisms by which compounds exert their biological effects.
Highlights
Introduction), androgen receptor (AR), glucocorticoid receptor (GR), mineralocorticoid receptor (MR) and progesterone receptor (PR) [1]
Steroid hormone receptors are ligand-inducible transcription factors belonging to nuclear hormone receptor superfamilly and include two receptors for estrogens (ER and ER), androgen receptor (AR), glucocorticoid receptor (GR), mineralocorticoid receptor (MR) and progesterone receptor (PR) [1]
The main advantage of the whole system is that it is based on single cell type: osteosarcoma U2OS cell line, and it has unified and constant cellular background that is defined by the expression of the same type and amounts of enzymes involved in the metabolism of compounds and the same coactivators and corepressors regulating the steroid receptor transcription response
Summary
), androgen receptor (AR), glucocorticoid receptor (GR), mineralocorticoid receptor (MR) and progesterone receptor (PR) [1]. They control essential physiological, developmental, reproductive and metabolic processes. In addition to their indisputable numerous roles in the physiology of healthy organisms, a growing body of work indicates they represent an important pharmaceutical target in number of diseases including cancer. Emerging evidence for the tumor suppressive functions of ER in many cancers like breast [6], colon [7], ovarian [8] and prostate [9] cancer make the ER selective agonists promising drugs for the future treatment of these tumors
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