双向凝胶电泳结合激光捕获显微切割技术成功构建人正常前列腺细胞的双向电泳图谱

Two-dimensional Gel Electrophoresis Profiles of Human Normal Prostate Cells Successfully Established by 2-DE Coupled with Laser Capture Microdissection

目的:建立正常前列腺细胞的蛋白质组学研究的技术体系,全面展示正常前列腺细胞的蛋白质表达谱。方法:用激光捕获显微切割(LCM)技术获取正常前列腺组织中的上皮细胞,提取细胞中的蛋白质,建立固相pH梯度双向凝胶电泳图谱,应用图像扫描仪及ImageMaster 2D Evolution分析软件获得蛋白质点的数字化和匹配性信息,挑选匹配良好的高丰度蛋白点,进行基质辅助激光解析/电离飞行时间质谱(MALDI-TOF-MS)鉴定。结果:成功构建出正常前列腺细胞的双向电泳图谱,并用MALDI-TOF-MS成功鉴定出正常前列腺细胞的一个蛋白质点。结论:正常前列腺细胞的蛋白质样品双向电泳图谱重复性良好,初步建立了正常前列腺细胞的蛋白质组学研究技术并验证了该技术的可行性。

Objective： To establish a technological system of proteomics research for displaying the full-scale protein expression spec- trum of the prostate cells. Methods ： We obtained the epithelial cells from normal prostate tissues by laser capture microdissection （LCM） technology, extracted the proteins from them and established two-dimensional gel electrophoresis （2-DE） profiles of the pro- teins by immobilized pH gradient（IPG） two-dimensional electrophoresis. Then protein spot detection and matching were performed with the scanner and the ImageMaster 2D Elite ananlysis software. With the help of the Ettanspotter and digester, protein spots that matched well aross the gels were extracted, digested and further identified by assisted laser desorption/ionization time of flight mass spectrometry （MALDI-TOF-MS）. Results ： The 2-DE profiles of prostate cells were established, and one protein spot from normal prostate cells i- dentified successfully. Conclusion ： The 2-DE profiles of normal prostate cells have good reproducibility. We have tentatively estab- lished the proteomics research technology of normal prostate cells and demonstrated the feasibility of proteomics research with the model of human normal prostate cells. Natl J Androl, 2007, 13 （12） ：1064-1067