Department of Food Science and Biotechnology, Dongguk University, Goyang, 10326, Republic of Korea.
Department of Integrated Biomedical and Life Science, Korea University, Seoul, 02841, Republic of Korea.
Chem Biol Interact. 2021 Nov 1;349:109655. doi: 10.1016/j.cbi.2021.109655. Epub 2021 Sep 11.
To develop a novel cell-based assay to evaluate the androgenic endocrine-disrupting properties of chemical substances, we established a method to detect ligand-mediated androgen receptor (AR) dimerization in stably transfected human cell lines using a bioluminescence resonance energy transfer (BRET) system. Using stably transfected human embryonic kidney (HEK-293) cells, the BRET-based AR dimerization assay was optimized as a novel test method and was validated using test chemicals recommended by the Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM). The BRET-based AR dimerization assay showed high accuracy, sensitivity, and specificity for the detection of androgenic endocrine-disrupting chemicals (EDCs), and the assay protocol is adequate for practical use. This dimerization assay is based on ligand-mediated hormone receptor dimerization and can provide accurate information about androgenic endocrine-disrupting properties at the cellular level, complementing conventional binding and transactivation assays.
为了开发一种新型的基于细胞的测定方法来评估化学物质的雄激素内分泌干扰特性，我们建立了一种使用荧光共振能量转移（BRET）系统检测稳定转染的人细胞系中配体介导的雄激素受体（AR）二聚化的方法。使用稳定转染的人胚肾（HEK-293）细胞，基于 BRET 的 AR 二聚化测定法被优化为一种新型的测试方法，并使用替代方法验证协作委员会（ICCVAM）推荐的测试化学品进行了验证。基于 BRET 的 AR 二聚化测定法对雄激素内分泌干扰化学物质（EDCs）的检测具有高准确性、灵敏度和特异性，并且该测定协议足以实际使用。这种二聚化测定法基于配体介导的激素受体二聚化，可以在细胞水平上提供关于雄激素内分泌干扰特性的准确信息，补充了传统的结合和转激活测定法。
