Human cell-based estrogen receptor beta dimerization assay

Abstract

Estrogen is not only responsible for important functions in the human body, such as cell growth, reproduction, differentiation, and development, but it is also deeply related to pathological processes, such as cancer, metabolic and cardiovascular diseases, and neurodegeneration. Estrogens and other estrogenic compounds have tran-scriptional activities through binding with the estrogen receptor (ER) to induce ER dimerization. The two es-trogen receptor subtypes, estrogen receptor alpha (ER alpha) and estrogen receptor beta (ER8), show structural differences and have different expression ratios in specific cells and tissues. Currently, the methods for con-firming the estrogenic properties of compounds are the binding (Test guideline no. 493) and transactivation (Test guideline no. 455) assays provided by the Organization for Economic Co-operation and Development (OECD). In a previous study, we developed an ER?? dimerization assay based on the bioluminescence resonance energy transfer (BRET) system, but there are currently no available tests that can confirm the effect of estrogenic compounds on ER8. Therefore, in this study, we developed a BRET-based ER8 dimerization assay to confirm the estrogenic prosperities of compounds. The BRET-based ER8 dimerization assay was verified using nine repre-sentative ER ligands and the results were compared with the dimerization activity of ER??. In conclusion, our BRET-based ER8 dimerization assay can provide information on the ER8 dimerization potential of estrogenic compounds.