姜黄素逆转人结肠癌细胞HCT-8/VCR的双向凝胶电泳技术的建立及优化  

Establishment and Optimization of Two-dimensional Gel Electrophoresis for Multidrug-resistance Reversing of Curcumin on Human Colon Carcinoma HCT-8/VCR Cells

作　　者：李雷[1] 傅仲学[1] 覃勇[1] 卢伟东[1] 吴星烨[1] 汤为学[1] 

机构地区：[1]重庆医科大学附属第一医院外三科,重庆400016

出　　处：《世界科技研究与发展》2011年第6期1074-1077,共4页World Sci-Tech R&D

基　　金：重庆市卫生局中医药重点项目(2009-1-2)资助项目

摘　　要：目的建立并优化姜黄素逆转人结肠癌细胞HCT-8/VCR蛋白质组的双向电泳技术。方法姜黄素作用于人结肠癌细胞HCT-8/VCR,提取蛋白质。对双向凝胶电泳中蛋白质样品的处理,蛋白上样量,IPG胶条的选择,以及聚焦条件等进行调整和优化。结果采用瓶内刮取的方法裂解细胞,pH3-10NL的胶条,上样量200 ug,延长除盐时间,聚焦60 000伏小时,可以得到分辨率较高、重复性较好的双向电泳图谱。结论初步建立了分辨率较高且重复性较好的姜黄素逆转人结肠癌细胞HCT-8/VCR蛋白质双向凝胶电泳图谱。Objective To establish and optimize a two-dimensional electrophoresis method for multidrug-resistance reversing of cureumin on human colon carcinoma HCT-8/VCR cells. Methods After the cultured colon carcinoma HCT-8/VCR cells were treated by cureumin, the proteins were extracted. The method of deal with protein samples, protein quantities, the choice of immobilized pH gradient（IPG） strips, isoe- lectric focusing（IEF） conditions were used ＂for establishing two-dimensional electrophoresis. Results The best 2-D separation results could be done through loading 200 ug of protein lysate which is directly scraped off from the plates, using the non-linear IPG strip and longer isoelectric focusing runs to eliminate salts,and 60 000 Volts. Hours. Conclusion The well-resolved, reproducible 2-DE profiles of multidrug-rcsistance reversing of curcumin on human colon carcinoma HCT-8/VCR cells have been primarily established.

关 键 词：蛋白质组学 双向凝胶电泳 姜黄素 多药耐药 结肠癌 

分 类 号：R285[医药卫生—中药学]