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A Chemical Screening Procedure for Glucocorticoid Signaling with a Zebrafish Larva Luciferase Reporter System
Published on: September 10, 2013
An efficient and economic high-throughput cell screening model targeting the glucocorticoid receptor
Yanli Zhang1, Feng Guo, Chengyu Jiang
1National Laboratory of Medical Molecular Biology, Dept. of Biochemistry & Molecular Biology, Chinese Academy of Medical Sciences, Peking Union Medical College, No. 5 dongdansantiao, Beijing, Peoples Republic of China. zhyanli@gmail.com
Journal of Drug Targeting
|January 4, 2008
Summary
A new cell line efficiently detects glucocorticoid receptor (GR) activation, enabling high-throughput screening for drugs targeting inflammatory diseases like rheumatoid arthritis and asthma.
Area of Science:
- Molecular Biology
- Cell Biology
- Pharmacology
Background:
- Glucocorticoid receptor (GR) activation is crucial for treating inflammatory diseases.
- Developing efficient screening methods is essential for discovering novel GR-targeting therapeutics.
Purpose of the Study:
- To establish and evaluate a high-throughput drug screening cell model for identifying compounds that activate the glucocorticoid receptor (GR).
- The model utilizes green fluorescent protein 4 (GFP4) as a marker for GR activation, aiming to improve treatments for inflammatory conditions.
Main Methods:
- A cell line (293E/GRE8x/GFP4) was engineered by stably transfecting 293E cells with a plasmid containing eight copies of the glucocorticoid response element (GRE) linked to GFP4.
- Selection and validation involved dexamethasone (DEX) induction and phorbol myristate acetate (PMA) inhibition, assessed via fluorescence microscopy and flow cytometry.
Main Results:
- The GFP4 expression in the engineered cell line was confirmed to be GRE-controlled, dose-dependently upregulated by DEX (EC50 ≈ 5 x 10⁻⁸ M), and downregulated by PMA (IC50 ≈ 3 x 10⁻⁶ gl⁻¹).
- Triamcinolone acetonide effectively activated GFP4 expression, demonstrating the cell line's responsiveness to GR agonists.
Conclusions:
- The developed 293E/GRE8x/GFP4 cell line serves as a sensitive and effective platform for high-throughput drug screening.
- This model is suitable for identifying novel therapeutic agents targeting the GR for the treatment of inflammatory diseases.
