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

目的建立并优化姜黄素逆转人结肠癌细胞HCT-8／VCR蛋白质纽的双向电泳技术。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每化。结果采用瓶内刮取的方法裂解细胞，pH3-10NL的胶条，上样量200ug，延长除盐时间，聚焦60000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。

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

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.