GPER Mediates Non-Genomic Effects of Estrogen

Estrogens are important modulators of a broad spectrum of physiological functions in humans. However, despite their beneficial actions, a number of lines of evidence correlate the sustained exposure to exogenous estrogen with increased risk of the onset of various cancers. Mainly these steroid hormo...

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Veröffentlicht in:Methods in molecular biology (Clifton, N.J.) Jg. 1366; S. 471
Hauptverfasser: Pupo, Marco, Maggiolini, Marcello, Musti, Anna Maria
Format: Journal Article
Sprache:Englisch
Veröffentlicht: United States 2016
Schlagworte:
Breast Neoplasms - genetics
Breast Neoplasms - metabolism
Breast Neoplasms - pathology
Cell Line, Tumor
Cell Migration Assays
Cell Movement - drug effects
Chromatin Immunoprecipitation
Connective Tissue Growth Factor - genetics
Connective Tissue Growth Factor - metabolism
Early Growth Response Protein 1 - genetics
Early Growth Response Protein 1 - metabolism
Estradiol - pharmacology
Estrogens - pharmacology
Female
Fibroblasts - drug effects
Fibroblasts - metabolism
Fibroblasts - pathology
Genes, Reporter
Humans
Luciferases - genetics
Luciferases - metabolism
Mitogen-Activated Protein Kinases - metabolism
Phosphorylation
Promoter Regions, Genetic
Proto-Oncogene Proteins c-fos - genetics
Proto-Oncogene Proteins c-fos - metabolism
Receptors, Estrogen - genetics
Receptors, Estrogen - metabolism
Receptors, G-Protein-Coupled - agonists
Receptors, G-Protein-Coupled - genetics
Receptors, G-Protein-Coupled - metabolism
Signal Transduction - drug effects
Transfection
Workflow
Breast cancer
Non-genomic signaling
GPR30 /GPER
Estrogen
MAPK phosphorylation
ISSN:1940-6029, 1940-6029
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Zusammenfassung:Estrogens are important modulators of a broad spectrum of physiological functions in humans. However, despite their beneficial actions, a number of lines of evidence correlate the sustained exposure to exogenous estrogen with increased risk of the onset of various cancers. Mainly these steroid hormones induce their effects by binding and activating estrogen receptors (ERα and ERβ). These receptors belong to the family of ligand-regulated transcription factors, and upon activation they regulate the expression of different target genes by binding directly to specific DNA sequences. On the other hand, in recent years it has become clear that the G protein-coupled estrogen receptor 30 (GPR30/GPER) is able to mediate non-genomic action of estrogens in different cell contexts. In particular, GPER has been shown to specifically bind estrogens, and in turn to functionally cross-react with diverse cell signaling systems such as the epidermal growth factor receptor (EGFR) pathway, the Notch signaling pathway and the mitogen-activated protein kinases (MAPK) pathway. In this chapter we will present some of the different experimental techniques currently used to demonstrate the functional role of GPER in mediating non-genomic actions of estrogens, such as the dual luciferase assay, assessment of the involvement of GPER in the stimulation of cell migration in breast cancer cell lines and in cancer-associated fibroblasts, and chromatin immunoprecipitation assay. Overall, the experimental procedures described herein represent key instruments for assessing the biological role of GPER in mediating non-genomic signals of estrogen.
Bibliographie:ObjectType-Article-1
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content type line 23
ISSN:1940-6029
1940-6029
DOI:10.1007/978-1-4939-3127-9_37