采用蛋白指纹图谱技术筛选大肠癌肝转移特异性相关蛋白  

Specific serum protein of hepatic metastasis from colorectal cancer screened by fingerprint

作　　者：崔丹瑜[1] 姜泊[1] 王继德[1] 李霆[1] 吴保平[1] 

机构地区：[1]南方医科大学南方医院消化内科,广东省广州市510515

出　　处：《中国组织工程研究与临床康复》2007年第9期1632-1635,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：目的:应用表面增强激光解析电离飞行时间质谱技术从大肠癌及大肠癌肝转移患者中筛选出大肠癌肝转移患者血清特异性相关蛋白。方法:实验于2005-07/2006-09分别在南方医院消化中心实验室与解放军第一五○医院实验室完成。应用美国CipherGen公司IMAC3(ImmobilizedMetalAffinityCapture,金属亲和表面)芯片和蛋白芯片仪检测44例大肠癌患者及36例大肠癌肝转移患者血清中的蛋白质相对含量。设定所有血清样本检测的蛋白质相对分子质量区间在1500～20000。利用PBSⅡ型蛋白质芯片阅读仪对IMAC3芯片进行检测,所得到的蛋白质以波谱的形式表示。采用BiomarkerWizard软件对2组血清在相同质荷比的蛋白质含量数据进行方差分析,将分析所得到的含量有显著性差异(P<0.05)的蛋白质建立数据库,导入BiomarkerPattern智能统计分析软件,选择相应条件,对其进行分组统计,从而得到能够正确分组的特异性蛋白标志物并构建大肠癌肝转移的诊断模型。采用酶联免疫法检测相同血清标本中的CEA水平,与构建的诊断模型在大肠癌肝转移诊断中作比较。结果:①44例大肠癌患者与36例大肠癌肝转移患者的血清蛋白质在质荷比为2685.64～11813间有16个蛋白质含量有显著差异。②大肠癌组在质荷比为5909处的蛋白质的相对含量高于大肠癌肝转移组[(30.1±9.6)%,(14.5±10.4)%,P≤0.01]。③其中44例大肠癌患者中有38例患者被正确分组,36例大肠癌肝转移患者被正确识别,准确率为92.5%(74/80),灵敏度和特异性分别为100%(36/36),86.4%(38/44)。结论:表面增强激光解析电离飞行时间质谱技术快速、准确、灵敏度、特异性高,通过蛋白芯片仪发现的特异性相关蛋白,有望成为大肠癌肝转移诊断中有应用价值的临床检测指标。AIM： To discover special serum protein of hepatic metastasis in patients of colorectal cancer （CC） and hepatic metastasis of colorectal cancer （HMCC） by using the technology of surface enhanced laser desorption/ionization-time of flight-mass spectrometry （SELDI-TOF-MS）. METHODS： The experiment was completed in the 150 Hospital of Chinese PLA and the Digestion Central Laboratory of Nanfang Hospital from July 2005 to September 2006. We screened for potential tumor biomarkers of serum samples from 44 CC patients and 36 HMCC patients by using the technology of SELDI-TOF-MS and IMAC3 （Immobilized Metal Affinity Capture） Protein Chip （CipherGen Company, USA）. We defined the relative molecular mass of serum protein in the range from 1 500 to 20 000 Da. We used the PBS Ⅱ protein chip reader to analyze the IMAC3 protein chips and transform the protein information into the form of spectra. We analyzed protein content of two groups in the same mass-to-charge ratio （M/Z value） and proceeded the analysis of variance by Biomarker Wizard software. The proteins whose contents in serum were significantly different were selected to construct a protein DATA, which was divided into different groups and analyzed respectively. Then a diagnosis pattern of HMCC was established, which could distinguish the special protein biomarker correctly by Biomarker Pattern software. Meanwhile a comparison between this new diagnosis pattern of HMCC and classical method was made by testing the CEA level in serum of the same patients. RESULTS： （1）The contents of 16 proteins in the two groups were significantly different and the M/Z values of these 16 proteins were from 2 685.64 Da to 11 813 Da. （2）The relative protein content of CC group was higher than that of HMCC group at the M/Z value of 5 909 [（30.1±9.6）%, （14.5±10.4）%, P ≤ 0.01].（3）There were 38 in 44 CC patients and all the 36 HMCC patients were classified correctly. The ratio of correctness, sensitivity and specificity were 92.5% （

关 键 词：大肠癌 肝转移 生物标志物 蛋白指纹图谱 表面增强激光解析电离飞行时间质谱 

分 类 号：R318.6[医药卫生—生物医学工程]