Effect of ET-1, FGF-2, and HGF on intracellular steroidogenesis in prostate cancer cells.

Abstract

e16083 Background: In the prostatic epithelium endothelin-1 (ET-1), fibroblast growth factor-2 (FGF2) and hepatocyte growth factor (HGF) are involved in cell proliferation and inhibition of apoptosis. The aim of this study is to investigate whether neuropeptides such as ET-1 and growth factors such as FGF and HGF may constitute an alternate pathway to intracellular steroidogenesis. Methods: PC3 cells, an androgen independent prostate cancer cell line were cultured in RPMI 1640, fully supplemented with FBS 10% and in serum free conditions. The cells were incubated with ET-1(10nM, 3 hrs incubation), FGF-2 (10 ng/ml, 3 hrs incubation) and HGF(33 ng/ml, 3 hrs incubation), and then progesterone, 17-OH progesterone and aldosterone were measured in cell lysates by LC-MS using LTQ Orbitrap XL MS 2.5.5 SP1 (Thermo-Fisher) equipped with Accela AS 2.2.1, Accela pump. The data were analysed by Xcalibur 2.1.0 Software. Results: In baseline conditions PC3 cells produce no progesterone but high amounts of 17-OH progesterone and small amounts of aldosterone. Upon stimulation with endothelin there was a significant increase in progesterone, significant decrease in 17-OH progesterone and a very significant increase in aldosterone. Similarly stimulation with both HGF and FGF resulted in significant decreases in 17-OH progesterone and increases in aldosterone. Conclusions: Our results imply that in androgen deprivation conditions intracellular steroidogenesis can still take place by stimulation via seven membrane and tyrosine kinase receptors, contributing in part to a steroid dependent phenotype in addition to a steroid independent one, thus implying the potential for new therapeutic interventions.