应用差异凝胶电泳及质谱技术筛选肺癌患者血浆差异蛋白  

Screening of differentially expressed serum proteins of lung cancer by DIGE and MALDI-TOF MS/MS

作　　者：沙慧芳[1] 孙强玲[1] 杨晓华[1] 储天晴[2] 谢银银[3] 鲁静[3] 包国良[1] 冯久贤[1] 龚乐罗[1] 

机构地区：[1]上海交通大学胸科医院基础研究室,200030 [2]上海交通大学胸科医院.肺内科,200030 [3]上海交通大学胸科医院医学院瑞金医院血液学研究所,200025

出　　处：《上海交通大学学报（医学版）》2010年第4期399-403,共5页Journal of Shanghai Jiao tong University:Medical Science

基　　金：上海市市级医院新兴前沿技术联合攻关项目(SHDC12007103)~~

摘　　要：目的建立健康者、肺部炎症患者和肺癌患者的血浆蛋白表达谱,寻找与肺癌早期发生相关的差异蛋白。方法应用差异凝胶电泳(DIGE)和基质辅助激光解吸飞行时间质谱(MALDI-TOF-MS)对健康者、肺部炎症患者和Ⅰ期肺癌患者血浆进行蛋白质组学比较研究,寻找肺癌相关差异蛋白。结果应用DIGE进行分离后,成功获得了蛋白质组分辨率高、重复性好的双向凝胶电泳图谱;DeCyder 6.5软件分析获得差异大于2倍的蛋白质点共有72个;质谱鉴定去冗余后确定了12种蛋白质,其中7种与肿瘤相关,分别为酰基辅酶A合成酶家族成员3、抗血小板膜糖蛋白Ⅰb特异性抗体、γ-纤维蛋白原、4型血清淀粉样蛋白A、补体因子H、钙结合微丝蛋白、载脂蛋白Ⅰ。结论应用DIGE及MALDI-TOF-MS分离并初步鉴定了12种蛋白质,筛选出7种在肺癌患者血浆中高表达的肿瘤相关蛋白。Objective To establish plasma protein spectrum from healthy people, patients with pneumonia and patients with lung cancer, and seek differentially expressed proteins related to early development of lung cancer. Methods Differential gel electrophoresis （ DIGE） and matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry （ MALD1-TOF-MS） were employed to screen differentially expressed plasma proteins among healthy people, patients with pneumonia and patients with stage I lung cancer. Results Two dimensional gel electrophoresis patterns of plasma proteins with high-resolution and reproducibility were successfully obtained using DIGE. Seventy-two significantly differentially expressed protein spots were screened with DeCyder 6.5 software. Twelve proteins were successfully identified via peptide mass print using MALDI-TOF-MS, among which seven proteins were reported to be related to cancer, such as acyl-CoA synthetase family member 3, platelet membrane glycoprotein I b specific antibody, serum amyloid A4, gelsolin, complement factor H, fibrinogen gamma chain and apolipoprotein A- I . Conclusion Twelve proteins can be identified by DIGE and MALDI-TOF-MS, and 7 proteins highly expressed in plasma of patients with lung cancer can be screened.

关 键 词：差异凝胶电泳 蛋白组学 血浆 肺癌 

分 类 号：R734.2[医药卫生—肿瘤]