Rapid, non-destructive, cell-based screening assays for agents that modulate growth, death, and androgen receptor activation in prostate cancer cells.
BACKGROUND
We developed non-invasive, cell-based screening assays to rapidly and biologically assess factors that modulate prostate cancer growth and affect androgen receptor (AR) activity.


METHODS
LNCaP cells, which stably express enhanced green fluorescent protein (EGFP) either constitutively or upon AR activation, were treated with a variety of agents, and then monitored by fluorescence and MTS assays for dose-dependent changes in cell number and AR activity.


RESULTS
The assays were validated for rapid, fluorescence-based, quantitative measurement for the presence of growth and AR modulators. Using these assays, we found that osteoblast conditioned media (CM) enhanced prostate cancer cell growth, but not AR activity. After priming with androgen (<1 nM R1881), forskolin or the pesticide dichlorvos enhanced AR activation, whereas interleukin-6 (IL-6) inhibited it.


CONCLUSIONS
These non-destructive, cell-based assays enable rapid systematic monitoring of the effects of drugs or complex mixtures on prostate cancer cell growth and/or AR activity.
Tavassoli
Peyman
P
Department of Pathology & Laboratory Medicine, University of British Columbia, Vancouver, British Columbia, Canada.
Snoek
Rob
R
Ray
Mira
M
Rao
Leticia Gomez
LG
Rennie
Paul S
PS
D000230:Adenocarcinoma / Q000378:metabolism / Q000473:pathology*; D000728:Androgens; D017209:Apoptosis / Q000187:drug effects; D001189:Aryl Hydrocarbon Hydroxylases / Q000494:pharmacology; D018929:Cell Culture Techniques; D002455:Cell Division / Q000187:drug effects; D045744:Cell Line, Tumor; D005576:Colforsin / Q000494:pharmacology; D017077:Culture Media, Conditioned / Q000494:pharmacology; D000072467:Cytochrome P450 Family 2; D004006:Dichlorvos / Q000494:pharmacology; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D007306:Insecticides / Q000494:pharmacology; D015850:Interleukin-6 / Q000494:pharmacology; D008297:Male; D010006:Osteoblasts / Q000166:cytology; D011471:Prostatic Neoplasms / Q000378:metabolism / Q000473:pathology*; D011944:Receptors, Androgen / Q000378:metabolism; D039181:Steroid 16-alpha-Hydroxylase; D013250:Steroid Hydroxylases / Q000494:pharmacology
D000728:Androgens; D017077:Culture Media, Conditioned; D007306:Insecticides; D015850:Interleukin-6; D011944:Receptors, Androgen; D049452:Green Fluorescent Proteins; D005576:Colforsin; D004006:Dichlorvos; D013250:Steroid Hydroxylases; D001189:Aryl Hydrocarbon Hydroxylases; C450279:CYP2C11 protein, rat; D000072467:Cytochrome P450 Family 2; D039181:Steroid 16-alpha-Hydroxylase