ANDROGEN-REGULATED PROLIFERATION AND GENE-TRANSCRIPTION IN A PROSTATESMOOTH-MUSCLE CELL-LINE (PS-1)

Androgens play a key role in directing stromal-epithelial interactions in prostate gland development, in maintenance of adult phenotype, and in disease progression. To address the molecular mechanisms of androg en action in prostate stromal cells and the genes regulated by androge ns in this cell type, a stromal cell line (PS-1) was developed from ra t ventral prostate. The PS-1 cell line was adapted to chemically defin ed media and characterized as smooth muscle based on expression of des min, smooth muscle cu-actin, and nuclear androgen receptor, markers fo r prostate smooth muscle, To examine steroid hormone regulation, PS-1 cells were analyzed for response to a panel of steroid hormones in ser um-free, chemically defined media. PS-1 cells were significantly growt h stimulated by physiological con centrations of androgens (10 nM) rel ative to other steroids tested. To ascertain whether this cell line co uld be used to examine androgen-regulated gene expression, differentia l display PCR was used to demonstrate the presence of androgen-regulat ed transcripts and provide the initial steps in cloning these genes. R eplicate differential display PCR analyses showed consistent androgen stimulation of 16 messenger RNA species. Northern analysis confirmed a ndrogen regulation of 6 species. Sequence analysis of each indicated n o regions of significant homology or direct matches to existing sequen ces. Further study of the PS-I stromal cell line and androgen-regulate d genes identified here will provide a novel in vitro system for defin ing molecular mechanisms of androgen action in prostate stromal cells and the significance of stromal androgen-regulated genes to stromal-ep ithelial interactions.