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目的分析利用蛋白质组学方法研究胃癌耐药相关蛋白质中双向电泳凝胶的染色显示。方法培养胃癌细胞SGC7901和长春新碱诱导的耐药胃癌细胞SGC7901/VCR,用双向凝胶电泳技术分离总蛋白,银染及胶体考马斯亮蓝染色,Image Scanner扫描仪扫描凝胶。结果获得了背景清晰、重复性好的双向凝胶电泳图谱,两种染色凝胶相比,硝酸银染色在样品少时显示更佳,过量则影响图像质量,而胶体考马斯亮蓝染色在上样量增加时的凝胶蛋白质点数目及丰度均增加,并无明显拖尾。结论两种显色方法受样品量影响较大,恰当选用有利于通过蛋白质组学研究胃癌耐药机制工作的进一步开展。
Objective To analyze the expression of two-dimensional electrophoresis gels in gastric cancer drug-resistance-related proteins using proteomics methods. Methods SGC7901/VCR cells were cultured in gastric cancer cells SGC7901 and vincristine, and total protein was isolated by two-dimensional gel electrophoresis. The silver staining and colloidal Coomassie brilliant blue staining were performed. Image Scanner scanner was used to scan the gel. Results Two-dimensional gel electrophoresis patterns with clear background and good reproducibility were obtained. Compared with the two stained gels, the silver nitrate staining showed better when there were few samples, and the excess affected the image quality, while the colloid Coomassie brilliant blue stained the sample volume. The number and abundance of gel protein spots increased as they increased, and there was no significant tailing. Conclusion The two chromogenic methods are greatly influenced by the sample size, and the appropriate selection is conducive to the further development of the study of gastric cancer drug resistance mechanism through proteomics.