The G Protein-Coupled Estrogen Receptor Agonist G-1 Inhibits Nuclear Estrogen Receptor Activity and Stimulates Novel Phosphoproteomic Signatures

Autor: P. Lee Ferguson, L. Cody Smith, Kimberly J. Ralston-Hooper, Tara Sabo-Attwood
Rok vydání: 2016
Předmět:
Proteomics
0301 basic medicine
Time Factors
Antineoplastic Agents
Hormonal
Diarylpropionitrile
Estrogen receptor
Breast Neoplasms
Cyclopentanes
Biology
Toxicology
Receptors
G-Protein-Coupled
03 medical and health sciences
Estrogen-related receptor alpha
chemistry.chemical_compound
0302 clinical medicine
Phenols
Tandem Mass Spectrometry
Nitriles
Humans
Benzhydryl Compounds
Estrogen receptor activity
Chromatography
High Pressure Liquid
Estrogen receptor beta
Cell Proliferation
Dose-Response Relationship
Drug
Estradiol
GPCR-Estrogen Receptor Agonist Acts as Inhibitor at Nuclear Estrogen Receptor
Phosphoproteins
Genistein
Molecular biology
Cell biology
030104 developmental biology
Receptors
Estrogen
chemistry
030220 oncology & carcinogenesis
MCF-7 Cells
Quinolines
Pyrazoles
Estrogen-related receptor gamma
Propionates
Estrogen receptor alpha
GPER
Signal Transduction
Zdroj: Toxicological Sciences. 151:434-446
ISSN: 1096-0929
1096-6080
DOI: 10.1093/toxsci/kfw057
Popis: Estrogen exerts cellular effects through both nuclear (ESR1 and ESR2) and membrane-bound estrogen receptors (G-protein coupled estrogen receptor, GPER); however, it is unclear if they act independently or engage in crosstalk to influence hormonal responses. To investigate each receptor’s role in proliferation, transcriptional activation, and protein phosphorylation in breast cancer cells (MCF-7), we employed selective agonists for ESR1 propyl-pyrazole-triol (PPT), ESR2 diarylpropionitrile (DPN), and GPER (G-1) and also determined the impact of xenoestrogens bisphenol-A (BPA) and genistein on these effects. As anticipated, 17β-estradiol (E2), PPT, DPN, BPA, and genistein each enhanced proliferation and activation of an ERE-driven reporter gene whereas G-1 had no significant impact. However, G-1 significantly reduced E2-, PPT-, DPN-, BPA-, and genistein-induced proliferation and ERE activation at doses greater than 500 nM indicating that G-1 mediated inhibition is not ESR isotype specific. As membrane receptors initiate cascades of phosphorylation events, we performed a global phosphoproteomic analysis on cells exposed to E2 or G-1 to identify potential targets of receptor crosstalk via downstream protein phosphorylation targets. Of the 211 phosphorylated proteins identified, 40 and 13 phosphoproteins were specifically modified by E2 and G-1, respectively. Subnetwork enrichment analysis revealed several processes related to cell cycle were specifically enriched by G-1 compared with E2. Further there existed a number of newly identified proteins that were specifically phosphorylated by G-1. These phosphorylation networks highlight specific proteins that may modulate the inhibitory effects of G-1 and suggest a novel role for interference with nuclear receptor activity driven by E2 and xenoestrogens.
Databáze: OpenAIRE
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