Recent findings have shown that, in addition to the genomic action of
steroids, through intracellular receptors, short-time effects could be mediated through binding to membrane sites. In the present study of
prostate cancer LNCaP cells, we report that
dihydrotestosterone and the non-internalizable analog
testosterone-BSA increase rapidly the release of
prostate-specific antigen (PSA) in the culture medium. Membrane
testosterone binding sites were identified through
ligand binding on membrane preparations, flow cytometry, and confocal
laser microscopy of the non-internalizable fluorescent analog
testosterone-BSA-
FITC, on whole cells. Binding on these sites is time- and concentration-dependent and specific for
testosterone, presenting a KD of 10.9 nM and a number of 144 sites/mg
protein (approximately 13000 sites/cell). Membrane sites differ immunologically for intracellular
androgen receptors. The secretion of PSA after membrane
testosterone receptor stimulation was inhibited after pretreatment with the actin cytoskeleton disrupting agent
cytochalasin B. In addition, membrane
testosterone binding modifies the intracellular dynamic equilibrium of monomeric to filamentous actin and remodels profoundly the actin cytoskeleton organization. These results are discussed in the context of a possible involvement of these sites in
cancer chemotherapy.