Mönkkönen Kati S, Aflatoonian Reza, Lee Kai-Fai, Yeung William S B, Tsao Sai-Wah, Laitinen Jarmo T, Fazeli Alireza
Department of Pharmacology and Toxicology, University of Kuopio, FIN-70211 Kuopio, Finland.
Mol Hum Reprod. 2007 Dec;13(12):845-51. doi: 10.1093/molehr/gam075. Epub 2007 Oct 31.
Heterotrimeric G proteins play a key role in membrane-mediated cell-signalling and hormonal regulation. Our earlier studies gave evidence of G protein subunit Galpha(i2) being under hormonal regulation in human in vivo. In this study, we used immortalized human oviduct epithelial cell line OE-E6/E7 as a model to study the hormonal regulation of Galpha(i2). We aimed at clarifying whether estradiol or progesterone could individually regulate the expression of Galpha(i2) and its potential signalling partners. Furthermore, we aimed to investigate which sex hormone receptors could potentially mediate the gene regulation in OE-E6/E7 cell line. OE-E6/E7 cells were cultured for 5 days with different concentrations of estradiol or progesterone. Quantitative real-time polymerase chain reaction (Q-PCR) was performed using cDNA of the hormone-treated cells to reveal any changes in gene expression. The presence of potential receptor targets in these cells was studied using PCR. Our data clearly showed that low concentrations of estradiol up-regulated the expression of Galpha(i2) gene and down-regulated the expression of membrane progesterone receptor mPRalpha gene in OE-E6/E7 cell line. Progesterone had no significant effect on Galpha(i2) gene expression, but it caused up-regulation of mPRalpha gene expression. In conclusion, it appears that sex hormones regulate the expression of Galpha(i2) and mPRalpha genes in a reverse manner in OE-E6/E7 cells. Our results suggest that estrogen receptor ERbeta mediates the regulatory effects of estradiol in these cells.
异源三聚体G蛋白在膜介导的细胞信号传导和激素调节中起关键作用。我们早期的研究证明了G蛋白亚基Gα(i2)在人体内受到激素调节。在本研究中,我们使用永生化人输卵管上皮细胞系OE-E6/E7作为模型来研究Gα(i2)的激素调节。我们旨在阐明雌二醇或孕酮是否能单独调节Gα(i2)及其潜在信号伴侣的表达。此外,我们旨在研究哪些性激素受体可能介导OE-E6/E7细胞系中的基因调节。将OE-E6/E7细胞用不同浓度的雌二醇或孕酮培养5天。使用激素处理细胞的cDNA进行定量实时聚合酶链反应(Q-PCR),以揭示基因表达的任何变化。使用PCR研究这些细胞中潜在受体靶点的存在。我们的数据清楚地表明,低浓度的雌二醇上调了OE-E6/E7细胞系中Gα(i2)基因的表达,并下调了膜孕酮受体mPRα基因的表达。孕酮对Gα(i2)基因表达没有显著影响,但它导致mPRα基因表达上调。总之,性激素似乎以相反的方式调节OE-E6/E7细胞中Gα(i2)和mPRα基因的表达。我们的结果表明,雌激素受体ERβ介导了雌二醇在这些细胞中的调节作用。