Expression and regulation of the estrogen receptors in PC-3 human prostate cancer cells

• Published in: Steroids 2016-03, Vol.107, p.74-86
• Main Authors: Pisolato, R., Lombardi, A.P.G., Vicente, C.M., Lucas, T.F.G., Lazari, M.F.M., Porto, C.S.
• Format: Article
• Language: English
• Subjects: Cell Line, Tumor; ER-α36; ERK1/2; Estradiol - pharmacology; Estrogen Receptor alpha - biosynthesis; Estrogen Receptor alpha - genetics; Estrogen Receptor beta - biosynthesis; Estrogen Receptor beta - genetics; Estrogen receptors α and β; Gene Expression Regulation, Neoplastic - drug effects; GPER; Humans; Male; MAP Kinase Signaling System - drug effects; MAP Kinase Signaling System - genetics; Neoplasm Proteins - biosynthesis; Neoplasm Proteins - genetics; PC-3 cells; Prostatic Neoplasms - genetics; Prostatic Neoplasms - metabolism
• ISSN: 0039-128X; 1878-5867
• DOI: 10.1016/j.steroids.2015.12.021

•There is a crosstalk between the ERα and ERβ to regulate the expression of each other in PC-3 human prostate cancer cells.•ER-α36 expression is regulated differently from ERα in PC-3 human prostate cancer cells.•ER-α36 may be involved in the ERK1/2 activity in the 17β-estradiol rapid action in PC-3 human prostate cancer cells. The aim of this study was to identify the expression, cellular localization and regulation of classic estrogen receptors ERα and ERβ, ER-α36 isoform and GPER in the androgen-independent prostate cancer cell line PC-3. In addition, we evaluated the relative contribution of these receptors to the activation of the ERK1/2 (extracellular signal-regulated protein kinases) signaling pathway. These four estrogen receptors were detected by Western blot assays and were shown by immunofluorescence assays to localize preferentially in extranuclear regions of PC-3 cells. In addition, treatment with 17β-estradiol (E2) (1μM) for 24h led to down-regulation of the classic estrogen receptors, whereas E2 at physiological concentration (0.1nM) for 24h tended to increase the levels of ERα and ERβ. Furthermore, the ERα-selective agonist PPT selectively increased the expression of ERβ and the ERβ-selective agonist DPN increased ERα levels. None of these treatments affected expression of the ER-α36 isoform. The unusual cytoplasmic localization of the classic estrogen receptors in these cells differs from the nuclear localization in the majority of estrogen target cells and suggests that rapid signaling pathways may be preferentially activated. In fact, treatment with selective agonists of ERα, ERβ and GPER induced ERK1/2 phosphorylation that was blocked by the respective antagonists. On the other hand, activation of ERK1/2 induced by E2 may involve additional mechanisms because it was not blocked by the three antagonists. Taken together, the results indicate that there is a crosstalk between ERα and ERβ to regulate the expression of each other, and suggest the involvement of other receptors, such as ER-α36, in the rapid ERK1/2 activation by E2. The identification of new isoforms of ERs, regulation of the receptors and signaling pathways is important to develop new therapeutic strategies for the castration-resistant prostate cancer.