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基于基因组编辑糖皮质激素敲除人前列腺癌细胞系的雄激素转录激活检测的增强。

Enhancement of androgen transcriptional activation assay based on genome edited glucocorticoid knock out human prostate cancer cell line.

机构信息

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.

出版信息

Environ Res. 2019 Apr;171:437-443. doi: 10.1016/j.envres.2019.01.027. Epub 2019 Jan 11.

Abstract

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.

摘要

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。

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