散发性大肠癌血清多肽肿瘤标志物的高通量筛选

High-throughput Screening for Tumor Peptide Markers of Sporadic Colorectal Cancer in Serum

目的:以常规体检者和明确诊断的大肠癌患者为研究对象,对其血清进行多肽谱分析,统计分析获得大肠癌特异血清多肽峰,为大肠癌的分子诊断提供理论依据,提高大肠癌的早期诊断水平。方法:1)收集研究对象外周非抗凝血并记录其人口学特征,将非抗凝血进行离心分离血清并保存;2)用Dynabeads RPC18磁珠分离提取血清蛋白质,Bruker UltraFlex TOF/TOF采集信号并用分析软件Clinprot tools 2.2(Bruker)分析筛选出大肠癌血清显著差异峰;3)用SPSS13.0分析大肠癌患者和健康人多肽峰的差异,进行Logistic回归分析差异多肽对形成大肠癌的影响。结果:本次研究共获得111名健康人和94名大肠癌患者的血清多肽峰信息,其中109名健康人和91名大肠癌患者同时具有性别、年龄等人口学信息。筛选出差异多肽峰105个,其中76个多肽在大肠癌患者和健康人间的分布差异有统计学意义(P<0.05)。运用Logistic回归分析,进入回归方程(P<0.05)的有:年龄,质荷比(m/z)分别为1061.10、1213.09、1607.32、1867.02、1897.95、2011.67和5078.81的七种多肽。结论:液体蛋白芯片飞行时间质谱系统可高效、精确地筛查血清多肽。大肠癌患者与健康人的血清多肽存在差异,筛选得到的质荷比(m/z)分别为1061.10、1213.09、1607.32、1867.02、1897.95、2011.67和5078.81的七种多肽可能作为早期诊断大肠癌的潜在肿瘤标志物。

Objective： This research was designed to screen for potential tumor peptide markers in serum in order to provide some evidences for early diagnosis, follow-up and therapy evaluation of colorectal cancer. Methods： 1. Fasting blood samples were collected. Plasma was separated and stored. 2.Plasma was pretreated with magnetic beads. Signals of serum peptides were collected by Bruker UltraFlex TOF/TOF. 3.Statistical analysis of the collected data to screen the peptides with statistical significance with Clinprot tools 2.2 （Bruker）and SPSS 13.0. Results： The serum peptide information of 111 healthy people and 94 patients was collected. 109 healthy people and 91 patients＇ information was complete. Of the 105 serum peptides with differential expressions between healthy and patient group, 76 peptides were statistically significant （P〈0.05）. Age and 7 of the peptide profiles （m/z 1061.10, 1213.09, 1607.32, 1867.02, 1897.95, 2011.67, 5078.81） were elected to the logistic regression equation （P〈0.05）. Conclusions： Magnetic bead-based sample fractionation method combined with MALDI-TOF-MS shows accurate and stable outcome. The results demonstrate that the screened 7 serum peptides are significantly correlated with occurrence of colorectal cancer and may serve as potential tumor markers for early diagnosis.