双向凝胶电泳及质谱分析法在筛选人胃癌标记物中的应用

目的初步筛选人胃癌相关抗原,为进一步认识胃癌的发生、发展机制及其早期诊断、靶向治疗提供依据。方法采用双向凝胶电泳及免疫印迹法分离人胃癌组织的总蛋白质,以PD Quest 8.0软件分析图谱;用基质辅助激光解析电离飞行时间质谱及蛋白质数据库搜索对差异蛋白点进行鉴定。结果获得分辨率高、匹配性好的人胃癌组织总蛋白双向凝胶电泳图谱,图像分析显示3块胶的点数为789±27、匹配点数为684±21、匹配率为86.7%;获得高分辨率的免疫印迹反应图谱,图像分析找出差异反应蛋白质19处,质谱鉴定共得到11种蛋白质(包括结蛋白、β微管蛋白、波形蛋白及胶转蛋白等)。结论利用血清蛋白质组分析可初步筛选、鉴定11种人胃癌相关抗原,此对进一步认识胃癌的发生、发展机制及其早期诊断、靶向治疗提供了依据。

Application of the human gastric cancer markers screening with two dimensional electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

Objective To screen human gastric carcinoma-associated antigens,so to further analysis the the occurrence and development mechanism,early diagnosis and targeting treatment of gastric cancer.Methods Two dimensional polyacrylamide gel electrophoresis(2-DE) technique and Western blot were used to separate the proteome from human gastric carcinoma tissue,the proteins hypothesized differently were identified through their Peptide Mass Fingerprints by PD Quest 8.0;the differentially expressed protein spots were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and protein database searching.Results The good 2-DE pattern including resolution and reproducibility was obtained,and the pattern analysis showed the average spots of 789±27,and 684±21 spots were matched,the average matching rate was 86.7%.Of the 19 differentially expressed proteins,11 were identified as human gastric carcinoma-associated antigens by MALDI-TOF-MS,including Desmin,Tublin,Vimentin,Transgelin,et al.Conclusion Eleven human gastric carcinoma-associated antigens can be found and identified by serologic proteome analysis,which provides useful information for analyzing the occurrence and development mechanism,early diagnosis and targeting treatment of human gastric cancer.