Intracellular Ca2+ regulation in a human prostate stromal cell culture

Abstract

Prostate stromal cell cultures are used in vitro to study the cellular pathophysiology of benign prostatic hyperplasia (BPH), but their functional properties are poorly understood. This study characterized intracellular Ca2+ ([Ca2+]i) regulation in a cultured cell line in comparison to freshly isolated cells, as a background to understanding contractile regulation and cellular proliferation in this tissue. Prostate stromal cells were isolated from either PrS6 cell cultures, with an extended life span by transfection with the SV40 T-antigen, tsA58-U19, or freshly obtained transition zone prostate samples, primary cells. [Ca2+]i was measured in vitro with the indicator Fura-2 by epifluorescence microscopy. Phenylephrine, high-K+, and caffeine induced Ca2+-transients in primary cells (resting [Ca2+]i 94 +/- 8 nM, n = 29; peak 193 +/- 26 nM, n = 19). In PrS6 cells resting [Ca2+]i was 96 +/- 8 nM (n = 78) and in 34 of these 78 cells, 30 microM phenylephrine increased [Ca2+]i to 296 +/- 28 nM. 5-methyl-urapidil (10-30 microM) inhibited this response in 10 of 16 cells. Spontaneous Ca2+-transients were also observed in 91% of phenylephrine-responsive cells, but in only 20% of non-responsive cells (P < 0.01). Ca2+-transients were also induced by high-K+ solution, and 20 mM caffeine. The latter abolished the response to subsequent phenylephrine application. Depletion of intracellular Ca2+ stores by caffeine or restoration from a Ca2+-free superfusate caused a substantial rise of [Ca2+]i. PrS6 prostate stromal cells express functional alpha1-adrenoceptors associated with spontaneous intracellular Ca2+-transients. They exhibit functional Ca2+ channels, intracellular Ca2+ stores, and Ca2+ entry induced by store depletion. Stromal cultures can therefore be used to characterize the cellular physiology of prostate stromal cell contraction and proliferation.