| 祝青青,金 杰,徐 敏,李记坤,靖大道.基于液相色谱-质谱技术的早期胃癌血清多肽组学研究[J].现代生物医学进展英文版,2018,(9):1616-1623. |
| 基于液相色谱-质谱技术的早期胃癌血清多肽组学研究 |
| The Serum Polypeptide Study of Early Gastric Cancer Based on Liquid Chromatography-mass Spectrometry |
| Received:December 19, 2017 Revised:January 23, 2018 |
| DOI：10.13241/j.cnki.pmb.2018.09.004 |
| 中文关键词: 早期胃癌 血清 液相色谱-质谱联用 多肽组学 |
| 英文关键词: Early gastric cancer Serum LC-MS/MS Peptidomics |
| 基金项目:上海市科学技术委员会基金项目（134119a3100） |
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| 中文摘要: |
| 摘要 目的：建立早期胃癌外周血清特征性多肽谱，分析其生物学特征，以探索一种特异且敏感的早期胃癌血清学诊断方法。方法：采集10例早期胃癌和10例正常对照血清，使用蛋白沉淀法去除高丰度血清蛋白质后，通过液相色谱-质谱联用技术重复三次进行多肽分离、离子化、质谱检测，将原始数据应用Label free方法中MaxQuant算法对肽段进行相对定量，分析两组差异多肽及差异多肽匹配蛋白。结果：EGC组和N组重复三次所得色谱图总体比较一致，多肽重复检出率分别为87.54%和85.67%。其中EGC组可重复检测到的Unique peptide有65条，匹配对应31个蛋白；在EGC组和N组血清中显著差异的Unique peptide有22条，匹配对应11个蛋白。对血清显著差异多肽所匹配蛋白进行生物学分析，发现这些蛋白质多数位于细胞外，功能类别主要涉及分子水平的序列变换、信号肽、N-糖基化位点等，信号通路主要富集在凝血级联通路和补体级联通路。结论：早期胃癌血清特异性多肽谱图的建立可望成为胃癌早期诊断的血清学标志物，后续试验将进一步针对这些差异多肽进一步分析验证，筛选出早期胃癌标志性多肽，同时应用大样本进行验证，筛选出特异高效的早期胃癌血清标志物。 |
| 英文摘要: |
| ABSTRACT Objective: To establish the specific serum polypeptide spectra and explore specific and sensitive serum markers for di- agnosis of early gastric cancer. Methods: Serum samples of 10 early gastric cancer cases and 10 normal cases were enrolled, using protein precipitation to remove out high abundance ratios serum protein. Mass spectrometry detection for peptides repeated three times by liquid chromatography tandem-mass spectrometry technology (LC-MS/MS), and applied MaxQuant algorithm of Label free method to quanti- tate peptides relatively, and analysed the differential polypeptides and the matching protein between the two groups. Results: Three re- peated mass spectrograms in EGC group and N group are consistent generally, and the detected polypeptide repeated rates are 87.54% and 85.67% respectively. There were 65 unique serum peptides that were detected in EGC group, matching with 31 proteins, and there were 22 differential unique serum peptides between the EGC group and N group, matching 11 proteins. Biological analysis of the serum different polypeptides matched proteins showed most of these proteins were found in extracelluar domain, and the functional categories mainly involved sequence variant, signal peptide, glycosylation site: N-linked, etc. The signal pathways were mainly enriched in the co- agualtion cascade pathway and the complement cascade pathway. Conclusion: Establishing the early gastric cancer serum specificity polypeptide spectra is expected to become serological markers in diagnosis of early gastric cancer. The following experiments will further analyse these differential peptides and screen early gastric cancer signal peptides, after verificating with large sample, then screen out spe- cific and effective serum markers for early gastric cancer. |
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