National Institute of Food and Drug Safety Evaluation, Osong 28159, Republic of Korea.
Department of Food Science and Biotechnology, Dongguk University, Goyang 10326, Republic of Korea.
Environ Res. 2019 Apr;171:437-443. doi: 10.1016/j.envres.2019.01.027. Epub 2019 Jan 11.
Endocrine-disrupting chemicals (EDCs) interfere with the biological activity of hormones. Among EDC's, (anti-)androgenic compounds potentially cause several androgen-related diseases. To improve the accuracy of an in vitro transactivation assay (TA) for detection of (anti-)androgenic compounds, We established the glucocorticoid receptor (GR) knockout 22Rv1/MMTV cell line by using an RNA-guided engineered nuclease (RGEN)-derived CRISPR/Cas system. The 22Rv1/MMTV GRKO cell line was characterized and validated by androgen receptor (AR)-mediated TA assay compared with the AR-TA assay using 22Rv1/MMTV. In conclusion, the AR-TA assay with the 22Rv1/MMTV GRKO cell line was more accurate, excluding the misleading signals derived from glucocorticoids or equivalent chemicals, and might be an effective method for screening potential (anti-)androgenic compounds.
内分泌干扰化学物质 (EDCs) 会干扰激素的生物活性。在这些 EDC 中，(抗)雄激素化合物可能会导致多种与雄激素相关的疾病。为了提高体外转录激活测定 (TA) 检测 (抗)雄激素化合物的准确性，我们使用 RNA 指导的工程核酸酶 (RGEN) 衍生的 CRISPR/Cas 系统建立了糖皮质激素受体 (GR) 敲除 22Rv1/MMTV 细胞系。通过与使用 22Rv1/MMTV 的雄激素受体 (AR) 介导的 TA 测定相比，对 22Rv1/MMTV GRKO 细胞系进行了特征描述和验证。总之，使用 22Rv1/MMTV GRKO 细胞系的 AR-TA 测定更准确，排除了来自糖皮质激素或等效化学物质的误导信号，并且可能是筛选潜在 (抗)雄激素化合物的有效方法。
