摘要
目的运用弱阳离子磁珠(magnetic beads based weak cation exchange,MB-WCX)联合基质辅助激光解吸离子飞行时间质谱(matrix assisted laser desorption ionization time of flight mass spectrometry,MALDI-TOF MS)建立结直肠癌血清蛋白组学诊断模型。方法收集我院正常对照(健康体检者)、结直肠癌术前及术后患者血清标本各72例,弱阳离子磁珠分离血清多肽,MALDI-TOF MS建立正常对照、结直肠癌术前及术后患者血清蛋白表达谱,Clin Prot Tools 2.0软件分析差异表达峰并建立诊断模型,液相色谱-电喷雾离子化质谱(liquid chromatography-eletronic spray ionization mass/mass,LC-ESIMS/MS)鉴定差异表达蛋白。结果对比分析正常对照、结直肠癌术前及术后血清蛋白图谱,共发现80个差异表达峰,12个峰差异具有统计学意义(均P<0.01),与对照组相比,其中9个差异峰在结直肠癌术前的血清蛋白图谱中显示升高,术后显示降低,3个峰在结肠癌术前的血清蛋白图谱中显示降低,术后显示升高。遗传算法(genetic algorithm,GA)模型诊断结直肠癌的敏感度和特异性分别为99.31%和96.49%。GA模型中m/z:2663.36、m/z:4793.17和m/z:5343.48的差异表达峰经鉴定分别为纤维蛋白原α前体亚型1(isoform 1 of Fibrinogen alpha chain precursor,FGA)、组蛋白赖氨酸甲基转移酶SETD7(histone-lysine N-methyltransferase SETD7,SETD7)和黏蛋白5AC(Mucin-5AC precursor,MUC5AC)。结论血清蛋白质谱模型能够准确区分正常对照与结直肠癌患者,但尚需更进一步研究证实。
Objective Serum protein expression profiling was examined using magnetic bead-based matrixassisted laser desorption/ionization time-of-flight（MALDI-TOF-MS） to establish a serum proteomic diagnostic model for colorectal cancer. Methods Serum samples of normal control（CRTL, n=72）, colorectal cancer（pre-operation CRC, n=72, and post-operation CRC, n=72） were collected from 2014-9-1 to 2016-9-1. Peptidome of all samples were extracted by magnetic-bead-based weak cation-exchange chromatography（MBWCX） and detected by calibrated Autoflex Ⅲ MALDI-TOF-MS. Peptide mass fingerprinting were analyzed by Clin Prot Tools 2.0 software, and the differentially-expressioned peptides were further identified using LCESI-MS/MS. Results MALDI-TOF-MS identified 80 peaks（m/z）, in which 12 peaks showed significant differences among CRTL, pre-operation and post-operation CRC patients（P〈0.01）. 9 peaks were up-regulated and 3 peaks were down-regulated in CRC compared with CRTL, and these peaks showed a tendency to CRTL after operation. Based on the GA model, CRC patients could be discriminated from CRTL with 99.31% sensitivity and 96.49% specificity. Moreover, 3 peaks（m/z： 2663.36, m/z： 4793.17 and m/z： 5343.48） of the GA model were identified as protein FGA, SETD7 and MUC5 AC respectively. Conclusion The serum proteomic diagnostic model could accurately distinguish between CRTL and CRC, but it needs further research.
作者
段宝军
白俊
郭彦峰
普彦淞
马国栋
DUAN Baojun;BAI Jun;GUO Yanfeng;PU Yansong;MA Guodong(Department of Oncology, Shaanxi Provincial People＇s Hospital, Xi ＇an 710068, China;Department of General Surgery, Xi＇an Central Hospital, Xi＇an 710003, China;Department of General Surgery, Shaanxi Provincial People＇s Hospital, Xi＇an 710068, China;Department of Pathology, Shaanxi Provincial People ＇s Hospital, Xi＇an 710068, China)
出处
《肿瘤防治研究》
CAS
CSCD
2018年第6期386-390,共5页
Cancer Research on Prevention and Treatment
关键词
结直肠癌
蛋白组学
诊断模型
Colorectal cancer
Proteomics
Diagnostic model
