Characterisation and validation of an in vitro transactivation assay based on the 22Rv1/MMTV_GR-KO cell line to detect human androgen receptor agonists and antagonists
| Autor: | Da-Woon Jung, Yooheon Park, Toine F.H. Bovee, Keith A. Houck, Patience Browne, Atsushi Ono, Yun Sook Kang, Masahiro Takeyoshi, Nathalie Delrue, Hee-Seok Lee, Miriam N. Jacobs, Anne Milcamps |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Agonist
Transcriptional Activation medicine.drug_class Drug Evaluation Preclinical Team Bioassays & Biosensors Toxicology 03 medical and health sciences Transactivation Gene Knockout Techniques Mice 0404 agricultural biotechnology Glucocorticoid receptor Receptors Glucocorticoid Cell Line Tumor Validation medicine Androgen Receptor Antagonists Animals Humans 030304 developmental biology OECD Test guideline 0303 health sciences Chemistry Antagonist Transcriptional activation assay Reproducibility of Results 04 agricultural and veterinary sciences General Medicine Transfection 040401 food science Molecular biology In vitro Androgen receptor Mammary Tumor Virus Mouse Cell culture Receptors Androgen Androgens Glucocorticoid Food Science medicine.drug |
| Zdroj: | Food and Chemical Toxicology, 152 Food and Chemical Toxicology 152 (2021) |
| ISSN: | 0278-6915 |
| Popis: | We describe the characterisation and validation of an androgen receptor (AR) transactivation assay for detection of AR agonists and antagonists using a stably transfected human prostate cancer cell line. This 22Rv1/mouse mammary tumour virus glucocorticoid knock-out cell line based AR transactivation assay was validated by criteria in Organisation for Economic Cooperation and Development Guidance Document 34 to determine if the assay performed equally well to the AR EcoScreen Assay included in Test Guideline for AR Transactivation (OECD TG 458). There was no Glucocorticoid Receptor (GR) crosstalk, and no changes in the AR DNA sequence in cells after the successful knock out of GR. Subsequently, the concordance of classifications of the 22 test chemicals was 100% in all laboratories. The AR agonistic and antagonistic inter-laboratory coefficients of variation based on log[10% effect for 10 nM DHT, PC10] and log[inhibitory response of 800 pM DHT by at 30%, IC30] from comprehensive tests were 2.75% and 2.44%, respectively. The AR agonist/antagonist test chemical classifications were consistent across AR EcoScreen ARTA assay data for 82/89%, and the balanced accuracy, sensitivity, and specificity were 83/90%, 88/100% and 78/80%, respectively. This assay was successfully validated and was approved for inclusion in TG 458 in 2020. |
| Databáze: | OpenAIRE |
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