Endocrine Abstracts

Androgen receptor (AR, NR3C4) is a 110-kDa ligand-activated transcriptional factor that belongs to the steroid hormone receptor superfamily. It has broad developmental effects and its mutations may have a great impact on pubertal development, genesis of prostatic hyperplasia, and even prostate cancer. Commonly known endogenous ligands of AR are testosterone and 5α-dihydrotestosterone (DHT). In the last years, several drugs and environmental pollutants were identified to alter AR activity leading to drug interactions or to endocrine disruption. Therefore, development of in-vitro experimental tools for analysing androgenic or antiandrogenic effects of various compounds is of great importance. In our work, we describe construction and characterisation of a novel stably transfected human reporter cell line AIZ-AR for assessment of transcriptional activity of human androgen receptor. Cell line AIZ-AR is derived from human prostate carcinoma epithelial cell line 22Rv1 that was stably transfected with a reporter plasmid containing three copies of androgen response regions (ARRs) followed by a single copy of androgen response element (ARE) from the promoter region of human prostate specific antigen (PSA) gene. Presented AIZ-AR cell line remained fully functional for over 60 days and more than 25 passages in the culture as well as after cryopreservation. Time-course analyses revealed that AIZ-AR cells allow detection of AR ligands as soon as after 8 h of the treatment. Upon dose–response analyses with 23 steroids in 96-well plate format, we observed induction of luciferase activity by androgens, but not by mineralocorticoids and oestrogens. Some glucocorticoids and progesterone also activated AR, but with potencies two to three orders of magnitude lower as compared with androgens. Taken together, we have developed a rapid, sensitive, selective, high-throughput and reproducible tool for detection of human AR ligands, with potential use in pharmacological and environmental applications.