蛋白质组学技术在环境内分泌干扰物分析上应用

Application of Proteomic Technology to Analysis of Endocrine Disruptors

为了探讨蛋白质组学技术在环境雌激素分析上的应用,用浓度为3.7×10-9mol/L的17β-雌二醇(E2)处理MCF-7细胞2或3天后进行双向电泳,对比不同的蛋白质提取、染色方法,利用光密度扫描仪将电泳图像数字化,并使用PDQuest软件进行图像分析,以观察雌激素诱导下的蛋白质表达图谱。结果表明,在蛋白质的提取上,冻融法比超声裂解法更有效;在蛋白质染色上,银染法比考马斯亮蓝染色法能得到更多可显示的蛋白质斑点。用E2处理72h后的MCF-7细胞,可检测到40个下调蛋白和5个上调蛋白。该蛋白质表达模式的改变提示了环境雌激素诱导下的特异性蛋白质表达图谱变化。本研究为分析雌激素类内分泌干扰物的内在作用机制提供了一种新的可行方法,也为建立其高通量筛选方法提供了新的思路。

This study was aimed to investigate environmental estrogens by proteomics technologies. The protein expression patterns were explored using 2-dimensional （2-D） gel electrophoresis. Estrogen inducing protein expression profiles were obtained by treating MCF-7 cell with 17 beta-estradiol （E2） at a concentration of 3.7xl0~mol/L for two or three days. The protocols for cytosol protein extraction and gel staining methods were evaluated. 2-D gels were scanned by Densitometer and image analysis was performed by PDQuest software （Bio-Rad）. Results showed that freeze-thaw method was much more efficient than sonication method in terms of protein extraction. Regarding gel staining, silver stain could achieve more visualized protein spots comparing with Coomassie Stain. It was observed that approximately 40 proteins were down- regulated and 5 proteins were up-regulated when MCF-7 cells were treated with E2 at 72h. The change of protein expression pattern has the great potential of prediction as it represents estrogen （E2 in this case） inducing protein expression profiles. The study provided a new clue to investigate the insight mechanism of estrogenic EDs and a new chance to screen EDs in the physiologically trustworthy method.