胃癌耐药研究中二维电泳凝胶显色方法的分析

Analysis for staining methods of two-dimentional patterns in drug resistant gastric carcinoma cells research

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 investigate staining methods for two-dimensional gel（2-DE） electrophoresis in muhidrug resistance of gastric cancer. Methods Cultured vincristine-resistant human gastric cancer cell line SGC7901/VCR and its parental cell line SGC7901. Variant amount protein of those cells were separated by 2- DE. Gels were stained with silver nitrate or colloidal Coomassie brilliant blue, and scanned by Image Scanner. Results Well-resolved, reproducible 2-DE patterns of SGC7901/VCR and SGC7901 were established. Silver staining was better when protein sample amount was low, overloaded protein will interfere resolution of the maps. Gels stained with colloidal Coomassie brilliant blue had more protein spots numbers and abundance without apparent trails when increased loading protein sample. Conclusion Two staining methods were influenced largely by the sum of protein samples, properly selection may be helpful for further study with proteomics in multidrug resistance of gastric cancer.