Article ID Journal Published Year Pages File Type
2593335 Reproductive Toxicology 2016 11 Pages PDF
Abstract

•Androgen receptor transcriptional activation (AR-TA) assay was developed and optimized using the 22Rv1/MMTV cell line.•22Rv1/MMTV cell line is stably transfected pMMTV-Luc plasmid in 22Rv1 cells contained endogenous AR.•In the pre-validation study, 20 compounds recommended by the ICCVAM were tested.•In the androgen agonist and antagonist test, 22Rv1/MMTV AR-TA assay results and ICCVAM classifications were 100% and 93% similar, respectively.

The endocrine-disrupting effects of androgenic signaling play crucial roles in several androgen-related diseases. In attempting to develop an in vitro cell line to be used in androgen receptor (AR)-mediated reporter gene assays, we developed a stable 22Rv1/MMTV cell line, which is a human prostate cancer cell line that endogenously expresses functional AR, to evaluate AR-mediated transcriptional activation (TA). Using 22Rv1/MMTV cells, we established and optimized a test protocol for the AR-TA assay and validated the proposed assay using 20 compounds recommended by the Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM). All the performance parameters for agonist and antagonist assays were 91–100% comparable between the 22Rv1/MMTV assay and the ICCVAM report. In conclusion, the AR-TA assay using 22Rv1/MMTV cells might be a quick and relatively inexpensive method for screening large numbers of chemicals for their potential to activate or inhibit AR-mediated gene transcription.

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