Watson C S, Campbell C H, Gametchu B
Human Biological Chemistry & Genetics Department, University of Texas Medical Branch, Galveston, TX 77555-0645, USA.
Exp Physiol. 1999 Nov;84(6):1013-22. doi: 10.1111/j.1469-445x.1999.01903.x.
Our laboratory has identified plasma membrane oestrogen receptors on a GH3/B6 rat pituitary tumour cell line and several sublines which produce rapid (within minutes), non-genomic responses to oestrogens. Oestrogen receptors have been identified by their binding to nine different antibodies (Abs) which together recognize at least seven epitopes on the oestrogen receptor-alpha. GH3/B6/F10 cells, a membrane oestrogen receptor-enriched subline, elevate intracellular calcium levels in response to 10 nM oestradiol. Prolactin release in these cells is triggered by both 1 pM and 1 nM oestradiol and diethylstilbestrol (DES). A membrane oestrogen receptor-alpha immunocytochemical signal rapidly disappears (at 3 min) and reappears (at 12-15 min) when 1 nM oestradiol, 10 nM diethylstilbestrol, or 10 nM nonylphenol is applied to the cells. This suggests that both oestrogens and xenoestrogens can utilize this alternative pathway for oestrogenic action. Xenoestrogens, which have so far shown weak effects in genomic assay systems, should now be retested for activity in eliciting membrane-initiated oestrogenic responses.
我们实验室已在GH3/B6大鼠垂体肿瘤细胞系及其几个亚系中鉴定出质膜雌激素受体，这些细胞系对雌激素产生快速（数分钟内）的非基因组反应。雌激素受体已通过其与九种不同抗体（Abs）的结合得以鉴定，这些抗体共同识别雌激素受体α上至少七个表位。GH3/B6/F10细胞是一种富含膜雌激素受体的亚系，对10 nM雌二醇有反应，可提高细胞内钙水平。这些细胞中的催乳素释放由1 pM和1 nM雌二醇以及己烯雌酚（DES）触发。当向细胞施加1 nM雌二醇、10 nM己烯雌酚或10 nM壬基酚时，膜雌激素受体α免疫细胞化学信号会迅速消失（3分钟时）并重新出现（12 - 15分钟时）。这表明雌激素和外源性雌激素都可以利用这种雌激素作用的替代途径。迄今为止，在外源基因组检测系统中显示出微弱作用的外源性雌激素，现在应该重新测试其引发膜启动的雌激素反应的活性。
