Prostate cancer cell growth inhibition by tamoxifen is associated with inhibition of protein kinase C and induction of p21waf1/cip1

Prostate cancer chemoprevention agents exhibit selective activity against early stage prostate cancer cells

“…17 Brie¯y, cells were ®xed in 70% ethanol at 4 C, overnight. Cells were then resuspended in PBS containing 50 kunit/ ml type III-A RNase (Sigma, St. Louis, MD) and incubated for 30 min at 37 C; DNA was then stained by the addition of propidium iodide.…”

“…17 Brie¯y, for each of eight individual cell lines tested, 2.5 ± 7.5610 5 cells were plated into 9 cm tissue culture dishes in the presence of the standard tissue culture media for individual cell lines. 24-h after plating, cells were extensively washed and cultured in serum free media (ie, standard media for individual cell lines, minus FBS and bovine pituitary extract).…”

“…13,17 Brie¯y, between 1200 and 2000 cells in a ®nal volume of 100 ml were added to each well of a 96-well plate and allowed to adhere overnight. Each of 10 different drugs, in 10 different serial dilution concentrations, were then added in a volume of 100 ml to each well.…”

“…After incubating for 3 days, thymidine incorporation was measured as described. 13,17 Cells were harvested onto glass ®ber ®lters (Packard, Meriden, CT) with a Packard cell harvester and counted in a Matrix 9600 Microplate counter (Packard). All microtiter assays in this study were run in replicates of six and were repeated at least once at a separate time.…”

“…17 Brie¯y, 24 h after plating cells in a 96-well format, cells were switched to serum free media as described above for TGFb assays. Cells were then treated with various concentrations of TGFb-1 for 24 h, after which thymidine incorporation was measured.…”