Estrogen receptors localization and signaling pathways in DU-145 human prostate cancer cells

Estrogen Receptors Promote Migration, Invasion and Colony Formation of the Androgen-Independent Prostate Cancer Cells PC-3 Through β-Catenin Pathway

“…The cells were wounded using 200 µl sterile pipette tips and then washed to remove detached cells and debris (28) and incubated in the absence (control, basal level of cellular function) and presence of 17β-estradiol (E2, 0.1 and 10 nM; Sigma Chemical Co.); ERβ-selective agonist DPN [10 nM; 2,3-bis(4-hydroxyphenyl)-propionitrile, Tocris Bioscience, Bristol, United Kingdom] and ERB-041 (10 nM; Sigma Chemical Co.) or ERα-selective agonist PPT [10 nM; 4,4' ,4"-(4-propyl-(1H)-pyrazole-1,3,5-triyl)trisphenol, Tocris Bioscience] for 24 h at 37 • C. The cells were also untreated or pretreated with ERβ-selective antagonist PHTPP [10 nM; 4-(2-phenyl-5,7-bis(trifluoromethyl)pyrazolo(1,5-a)pyrimidin-3-yl)phenol, Tocris Bioscience] or with a compound that disrupts the complex β-catenin-TCF/LEF transcription factor, PKF 118-310 (100 nM, Sigma-Aldrich Co) for 30 min. Incubation was continued in the absence and presence of DPN (10 nM) or ERB-041 (10 nM), for 24 h at 37 • C (18,21,22). The agonists and antagonists are highly selective, at these concentrations, as previously reported [(29, 30), (31), (20)].…”

“…The human androgen-independent prostate cancer cell line (PC-3) (derived from bone metastasis) was obtained from the American Type Culture Collection (Manassas, VA, United States). PC-3 cells cultures were carried out as previously described (18,(20)(21)(22).…”

“…These chambers were placed in 24-well plates containing culture medium with 10% FBS in the lower chamber. PC-3 cells in upper chambers were incubated in the absence (control) and presence of E2 (10 nM), DPN (10 nM) or PPT (10 nM) for 48 h at 37 • C. The cells were also untreated or pretreated with PHTPP (10 nM), MPP [10 nM; 1,3-bis(4-hydroxyphenyl)-4-methyl-5-(4-(2piperidinylethoxy)phenol)-1H-pyrazole dihydrochloride, Tocris Bioscience], PKF 118-310 (100 nM) (18,21,22) or VEGF specific inhibitor Bevacizumab (Avastin R , 25 ng) (35) for 30 min. Incubation was continued in the absence and presence of E2 (10 nM), DPN (10 nM) or PPT (10 nM), for 48 h at 37 • C. The membranes were washed thoroughly with 10 mM PBS, fixed in 4% paraformaldehyde for 30 min, and stained with 0.2% crystal violet for 10 min (28).…”

“…In androgen-independent prostate cancer cells PC-3 (20) and DU-145 (21), ERα and ERβ are mostly located in the extranuclear region of these cells. The activation of these receptors increases the phosphorylation of ERK1/2 (extracellular signal-regulated kinase1 and 2) and AKT (serine/threonine kinases) in PC-3 cells (18,20).…”

“…The activation of these receptors increases the phosphorylation of ERK1/2 (extracellular signal-regulated kinase1 and 2) and AKT (serine/threonine kinases) in PC-3 cells (18,20). On the other hand, phosphorylated AKT is not detected in DU-145 cells (21). The role of these pathways through activation of estrogen receptors (ERs) remains to be explored in both cells.…”