Abstract
Androgen receptor plays multiple physiological and pathological roles in human organism. In the current paper, we describe construction and characterization 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 transfected with reporter plasmid containing 3 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. AIZ-AR cells remained fully functional for more than 60 days and over 25 passages in the culture and even after cryopreservation. Time-course analyses showed that AIZ-AR cells allow detection of AR ligands as soon as after 8 hours of the treatment. We performed dose-response analyses with 23 steroids in 96-well plate format. We observed activation of AR by androgens, but not by estrogens and mineralocorticoids. Some glucocorticoids and progesterone also induced luciferase, but their potencies were 2-3 orders of magnitude weaker as compared to 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.
Highlights
Androgen receptor (AR, NR3C4) is a 110-kDa ligand-activated transcriptional factor that belongs to the steroid hormone receptor superfamily
We present a novel stably transfected human gene reporter cell line for assessment of AR transcriptional activity
AIZ-AR cell line is derived from human prostate carcinoma epithelial cell line 22Rv1 expressing endogenous AR that was transfected with reporter plasmid containing sequence of androgen response element from promoter of human prostate-specific antigen (PSA)
Summary
Androgen receptor (AR, NR3C4) is a 110-kDa ligand-activated transcriptional factor that belongs to the steroid hormone receptor superfamily. It has broad physiological functions, including developmental and psychological. AR is involved in several pathological situations, including genesis of prostatic hyperplasia and prostate cancer function or altered pubertal development due to its mutations [1]. In the absence of a ligand, AR primarily resides in the cytoplasm bound to chaperone proteins. AR translocates to the nucleus where it forms AR/AR homodimer, which binds specific DNA sequence known as androgen.
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