结肠癌蛋白质双向电泳技术的建立  

Establishment of two-dimensional electrophoresis methods for human colorectal cancer proteome

作　　者：马怡茗[1] 赵玫[1] 时岚[1] 罗清[1] 钟佳伶[1] 刘健[1] 王佳[1] 黄常志[1] 

机构地区：[1]中国医学科学院北京协和医学院肿瘤医院肿瘤研究所分子肿瘤国家重点实验室病因及癌变北京市重点实验室,北京100021

出　　处：《现代生物医学进展》2010年第9期1601-1603,共3页Progress in Modern Biomedicine

基　　金：国家重点基础研究发展规划项目(2007CB914700)

摘　　要：目的:建立结肠癌13cm和24cm非线性分离系统的2-D图谱,分析比较两者的分辨率。方法:提取结肠癌总蛋白,用pH3-10非线性干胶条对样品进行等电聚焦分离,并分别使用13cm和24cm电泳系统进行双向电泳,考马斯亮蓝G250染色,图像分析,比较对比两组2-D图谱,量化分析两种系统的分辨率差异。结果:在等点电3-10,分子量20-170kD范围内分别分离得到蛋白质斑点873个(13cm电泳系统)和1349个(24cm电泳系统)。对于24cm电泳系统,1mg蛋白质上样量的电泳图谱清晰,分辨率较好。结论:成功建立了高分辨率、简便易控的结肠癌蛋白质组双向电泳技术平台。Objective：To establish two-dimensional electrophoresis methods for human colorectal cancer proteome with 13 cm and 24 cm IPG strip.Methods：The total proteins of human colorectal cancer tissues were extracted by homogenization and separated by 13 cm or 24 cm immobilized pH gradient（IPG） strips prior to SDS polyacrylamide gel electrophoresis.The gels were stained according to a modified Neuhoff＇s cooloidal coomassie blue G-250 staining method.To evaluate the spot pattern of each gel,MagicScan software on imagescanner was used to scan the 2-D gels and the software of PDQuest system was employed for the image analysis.One of the samely expressed protein spots were analyzed by MALDI-TOF-MS.For PMF identification,the Mascot search engine was employed against the NCBInr protein database with a mass tolerance of ±50 ppm.Results：873 protein spots were detected in 13 cm system and 1349 protein spots were detected in 24 cm system.for protein identified,MASCOT scores for protein matching were 74 and 142 respectively.Conclusion：The two-dimensional electrophoresis profiles of human colorectal cancer proteome were obtained with high resolution.

关 键 词：蛋白质组学 双向电泳 结肠癌 

分 类 号：R735.35[医药卫生—肿瘤] Q51[医药卫生—临床医学]