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Phytoestrogens Regulate mRNA and Protein Levels of Guanine Nucleotide-Binding Protein Beta-1 Subunit (GNB1) in MCF-7 Cells

机译：植物雌激素调节MCF-7细胞中鸟嘌呤核苷酸结合蛋白β-1亚基（GN1）的mRNA和蛋白质水平

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Phytoestrogens (PEs) are non-steroidal ligands which regulate the expression of number of estrogen receptor-dependent genes responsible for a variety of biological processes. Deciphering the molecular mechanism of action of these compounds is of great importance because it would increase our understanding of the role(s) these bioactive chemicals play in prevention and treatment of estrogen-based diseases. In this study, we applied suppression subtractive hybridization (SSH) to identify genes that are regulated by phytoestrogens through either the classic nuclear-based estrogen receptor or membrane-based estrogen receptor pathways. Suppression Subtractive Hybridization, using mRNA from genistein (GE) treated MCF-7 cells as testers, resulted in a significant increase in GNB1 mRNA expression levels as compared with 10 nM 17-β estradiol or the no treatment control. GNB1 mRNA expression was upregulated 2- to 5-fold following exposure to 100.0 nM genistein. Similarly, GNB1 protein expression was up regulated 12- to 14-fold. Genistein regulation of GNB1 was estrogen receptor-dependent, in the presence of the antiestrogen ICI-182,780, both GNB1 mRNA and protein expression were inhibited. Analysis of the GNB1 promoter using ChIP assay showed a phytoestrogen-dependent association of estrogen receptor α (ERα) and β (ERβ) to the GNB1 promoter. This association was specific for ERα since association was not observed when the cells were co-incubated with genistein and the ERα antagonist, ICI. Our data demonstrate that the levels of G-protein, beta-1 subunit are regulated by phytoestrogens through an estrogen receptor pathway and further suggest that phytoestrogens may control the ratio of α-subunit to β/γ-subunits of the G-protein complex in cells.

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