利用2DE-MS技术筛选和鉴定大肠癌血清标志物  

Screening and identification of serum markers of colorectal cancer by two-dimensional electrophoresis mass spectrometry

作　　者：何渝军[1,2] 牟芝蓉[2] 黎万玲[2] 刘宝华[1] 吴玉章[2] 

机构地区：[1]第三军医大学大坪医院野战外科研究所普通外科,重庆400042 [2]第三军医大学基础部全军免疫学研究所,重庆400038

出　　处：《第三军医大学学报》2012年第18期1835-1838,共4页Journal of Third Military Medical University

基　　金：重庆市自然科学基金(CSTC2011BB5032)~~

摘　　要：目的利用2DE-MS技术对大肠癌患者血清和正常人血清蛋白进行比较分析,筛选和鉴定差异蛋白。方法收集正常人(n=10)和大肠癌患者(n=15)术前血清,进行去除白蛋白和免疫球蛋白及脱盐浓缩预处理,应用2DE-PAGE分离处理后血清蛋白,比较两者的差异蛋白质点,采用MALDI-TOF分析鉴定差异蛋白质。结果血清经预处理后,可提高样品的上样量,2DE-PAGE图谱的蛋白质点数明显增加,水平条带明显减少。大肠癌患者血清中共筛选2个差异的蛋白点,经质谱鉴定为转甲状腺素蛋白(transthyretin,TTR)和细胞角蛋白1(cytokeratin-1,CK-1)。其中TTR在大肠癌患者血清中为低表达,而CK-1为高表达。大肠癌患者血清中TTR含量为(245.87±60.72)mg/L,明显低于大肠腺瘤组和正常血清组[分别为(299.53±67.91)、(311.31±67.01)mg/L,P<0.01]。结论 2DE-MS技术是筛选和鉴定疾病血清标志物的良好工具,TTR可能是大肠癌潜在的血清标志物。Objective To screen the differentially expressed proteins and peptides in the serum of patients with colorectal cancer （CRC） using two-dimensional electrophoresis mass spectrometry （2DE-MS）. Methods Serum samples were obtained from 15 CRC patients （CRC group） before operation and 10 healthy volunteers （control group）, and then pretreated for removing albumins and immunoglobulins and desahing. The differentially expressed protein spots were observed by comparing 2DE-PAGE gel maps between the CRC group and control group, and were identified by MALDI-TOF mass spectrometry. Results Compared with that of the serum samples without pretreatment, the volume of the pretreated serum samples increased, and more pro- tein spots were detected. The comparison of the 2DE-PAGE gel maps between the CRC group and control group showed two differentially expressed protein spots, which were identified as transthyretin （TFR） and cytokeratin- 1 （ CK-1 ） by MS. TFR was lowly expressed and CK-1 was highly expressed in the serum of the CRC group. The serum level of TFR in the CRC group （245.87 ± 60.72mg/L） was significantly lower than that in the colo- rectal adenoma group （299.53 ± 67.91 mg/L） and control group （311.31 ± 67.01 mg/L） （P 〈 0.01 ）. Conclusion 2DE-MS is a useful tool for screening and identifying disease serum markers. TrR may be a potential serum marker of CRC.

关 键 词：大肠癌 蛋白质组学 二维电泳 血清标志物 

分 类 号：R446[医药卫生—诊断学] R730.43[医药卫生—临床医学]