G Protein-Coupled Estrogen Receptor-Selective Ligands Modulate Endometrial Tumor Growth
Figure 3
Activation of PI3K by estrogen in Hec50 cells is mediated by GPER. Hec50 cells were transfected with a marker of PIP3 production, the PH domain of Akt fused to monomeric red fluorescent protein (mRFP) yielding the marker PH-RFP. (a) PH-RFP-transfected Hec50 cells were stimulated with the following ligands: 10 nM estrogen (17E2), 10 μM 17-estrogen (17E2), or 10 nM estrogen in the presence of the PI3K inhibitor LY294001 (10 μM, 20 min pre-incubation; 17E2 + LY), the EGFR inhibitor AG1478 (25 μM, 60 min pre-incubation; 17E2 + AG) or the metalloproteinase inhibitor GM6001 (10 μM, 30 min pre-incubation; 17E2 + GM). Unstimulated designates vehicle only. (b) PH-RFP-transfected Hec50 cells were stimulated for 15 min with the GPER-selective agonist G-1 or estrogen at the indicated concentrations in the absence or presence of the GPER-selective antagonists G15 and G36 (cells were pretreated 15 min with G15 or G36 prior to stimulation with E2 or G-1) or in the presence of 10 nM G-1 and the PI3K, EGFR or metalloproteinase inhibitors as in (a).
