title: Novel stably transfected human reporter cell line AIZ-AR as a tool for an assessment of human androgen receptor transcriptional activity.
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.
authors: Department of Cell Biology and Genetics, Faculty of Science, Palacky University, Olomouc, Czech Republic.
authors: Iveta
authors: I
authors: Bartonkova
authors: Department of Cell Biology and Genetics, Faculty of Science, Palacky University, Olomouc, Czech Republic.
authors: Aneta
authors: A
authors: Novotna
authors: Department of Cell Biology and Genetics, Faculty of Science, Palacky University, Olomouc, Czech Republic.
authors: Zdenek
authors: Z
authors: Dvorak
mesh_terms: D059002:Androgen Receptor Antagonists / Q000494:pharmacology; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D002934:Cinnamates / Q000494:pharmacology; D002999:Clone Cells; D015925:Cryopreservation; D013196:Dihydrotestosterone / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D017930:Genes, Reporter; D006801:Humans; D006921:Hygromycin B / Q000031:analogs & derivatives / Q000494:pharmacology; D008156:Luciferases / Q000378:metabolism; D011944:Receptors, Androgen / Q000235:genetics* / Q000378:metabolism; D013256:Steroids / Q000494:pharmacology; D013997:Time Factors; D014158:Transcription, Genetic / Q000187:drug effects; D014162:Transfection
chemical_list: C410798:AR protein, human; D059002:Androgen Receptor Antagonists; D002934:Cinnamates; D011944:Receptors, Androgen; D013256:Steroids; D013196:Dihydrotestosterone; D006921:Hygromycin B; C026273:hygromycin A; D008156:Luciferases