1. GPER Mediates Non-Genomic Effects of Estrogen.
- Author
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Pupo M, Maggiolini M, and Musti AM
- Subjects
- Breast Neoplasms genetics, 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, Estrogens pharmacology, Female, 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 genetics, Receptors, G-Protein-Coupled metabolism, Signal Transduction drug effects, Transfection, Workflow, Breast Neoplasms metabolism, Estradiol pharmacology, Fibroblasts drug effects, Receptors, G-Protein-Coupled agonists
- Abstract
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.
- Published
- 2016
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