联用2-DE和MALDI-TOF-MS技术筛查胃癌组织和血清中的差异蛋白质

目的 分别建立肠型、弥漫型胃癌组织和血清双向凝胶电泳图谱,鉴定差异表达蛋白,从中发现有意义的胃癌肿瘤相关标志物.方法 采用双向凝胶电泳（two-dimensional polyacrylamide gel electrophoresis,2-DE）分离组织和血清总蛋白,经硝酸银染色,分析选取有表达差异的蛋白质点,通过基质辅助激光解析电离飞行时间质谱（matrix-assisted laser desorption ionization time of flight mass spectrometry,MALDI-TOF-MS）鉴定差异蛋白.结果 获得重复性和分辨率较好的胃癌组织和血清双向凝胶电泳图谱,经MALDI-TOF-MS鉴定得到20种蛋白,在7例（58.3%）肠型胃癌组织中均有硒结合蛋白1（SBP1）表达,KIAA基因家族蛋白在弥漫型胃癌组织和血清中均有表达.结论 建立了弥漫型肠型胃癌组织和血清双向凝胶电泳图谱,获得了差异蛋白质点,为寻找胃癌的特异蛋白质作为胃癌肿瘤标志物奠定了基础.

2-DE combined with MALDI-TOF-MS technique in screening of differential proteins in tissue and serum of gastric cancer

Objective To identify the differential proteins expressed in gastric cancer. Methods Two-dimensional polyacrylamide gel electrophoresis （2-DE） was applied to separate proteins from tissue and serum samples of patients with gastric cancer. After silver staining, the differentially expressed proteins were analyzed by Imagemaster 6.0 software and identified by matrix-assisted laser desorption ionization time of flight mass spectrometry （MALDI-TOF-MS）. Results 2-DE profiles with high resolution and reproducibility were obtained. Twenty kinds of proteins were identified by MALDI-TOF-MS, among which 8 proteins were already reported. Protein SBP1 was found in 7 （58.3%） intestinal type gastric cancer and proteins of KIAA gene family were expressed in tissue and serum samples of diffuse gastric cancer. Conclusion The well-reproducible 2-DE profiles in tissue and serum of human gastric cancer were established. Differentially expressed proteins were obtained which provide experimental basis for screening cancer-related proteins of gastric cancer.