双向凝胶电泳进行肾癌与健康人血清中差异表达蛋白的筛选和鉴定  

Two-dimensional gel electrophoresis in screen and identification of differentially expressed proteins between patients with small renal cell carcinoma and healthy controls

作　　者：石农[1] 徐罡[2] 袁海川[1] 温晖[2] 顾斌[2] 刘俊[2] 马志承[2] 张立旻[2] 项翠琴[3] 陆晔[3] 丁强[2] 张元芳[2] 

机构地区：[1]复旦大学附属华山医院宝山仁和分院泌尿外科,上海200443 [2]复旦大学附属华山医院泌尿外科 [3]上海市疾病预防控制中心

出　　处：《上海医学》2011年第7期547-551,F0003,共6页Shanghai Medical Journal

基　　金：上海市卫生局青年基金(2009Y005);上海市宝山区科委项目(09-E-27);国家自然科学基金青年基金(81001130)项目资助

摘　　要：目的应用双向凝胶电泳(2-DE)结合基质辅助激光解析电离化/飞行时间质谱(MALDI-TOF-MS)技术对小肾癌与健康人的血清蛋白进行差异蛋白质的筛选和鉴定,以寻找潜在的肾癌肿瘤标志物。方法选取20例小肾癌患者和20名健康人,各抽取少量静脉血,分离出血清,应用2-DE结合MALDI-TOF-MS技术测定血清蛋白质表达谱,并用Mascot软件搜索NCBInr数据库,进行质谱学的鉴定。结果在凝胶图像上,共发现27个差异蛋白质位点。进一步对其中的5个位点进行切胶、酶解和质谱鉴定,分别为凝血因子ⅩⅢ、补体C3、C3前体、血液结合素和α1-B-糖蛋白。结论 2-DE技术鉴定出的5个在小肾癌患者与健康人血清中的差异表达蛋白质有可能在肾癌的早期诊断中起一定的肿瘤标志物的作用。Objective To identify the serum proteomic differences between patients with renal cell carcinoma （RCC） and healthy controls using two-dimensional gel electrophoresis and matrix assisted laser desorption ionization/time of flight MS（MALDI-TOF-MS）, in an effort to search for potential biomarkers for RCC. Methods We compared the serum protein profiles of 20 patients with small RCC with those of 20 healthy controls to identify the differentially expressed proteins. The serum protein expression profiles were analyzed by two- dimensional gel electrophoresis and MALDI-TOF-MS. Mascot software and NCBInr database were used. Results Twenty-seven differential protein sites were identified by two-dimensional gel electrophoresis between small RCC patients and healthy volunteers. Five differentially expressed proteins were chosen for further identification with mass spectrometry; they were coagulation factor XIII B, complement C3, C3 precursor, hemopexin, and alpha-l-B-glycoprotein. Conclusion We have identified five differential serum proteins between small RCC patients and healthy volunteers using 2-D gel electrophoresis; they may serve as novel biomarkers for early detection and diagnosis of human ROD.

关 键 词：肾细胞癌 血清标志物 双向凝胶电泳 

分 类 号：R737.11[医药卫生—肿瘤]