多维度CA19-9载体糖蛋白质组分析新方法及其在胰腺癌诊断中的应用

CA19-9是胰腺癌黄金标志物。临床使用中发现CA19-9定量容易造成假阳性结果。有报道表明单个CA19-9载体蛋白在胰腺癌诊断方面具有更大的潜力，但是由于CA19-9蛋白含量低、糖链结构复杂以及分析技术限制，尚缺少CA19-9糖链和糖肽的研究，因而无法判断蛋白是否真连有CA19-9糖链，更无法分辨CA19-9载体蛋白变化是糖链变化还是蛋白变化。本项目拟建立一种高灵敏的CA19-9载体糖蛋白分析策略。通过建立集成化样品前处理装置富集微量组织样品中的CA19-9糖肽，以及优化色谱质谱条件和解析糖链和糖肽结构，完成对CA19-9糖链和糖肽的定性定量分析。最后采用发展的方法鉴定胰腺癌的CA19-9糖肽和糖肽，并与其他非癌组织比较筛选出灵敏度高和特异性好的胰腺癌的标志物。采用该策略可以提高CA19-9的定量维度，项目的顺利实施将为胰腺癌的诊断提供新的思路。

CA19-9, as the gold standard marker of pancreatic cancer, easily leads to a false-positive result in clinical diagnosis due to the one-dimensional quantification. The CA19-9 carrier protein has been reported with greater potential in the diagnosis of pancreatic cancer. However, few studies focused on the comprehensive identification of CA19-9 glycans and glycopeptides because of the low abundance of CA19-9 carrier proteins, identification difficulties of entire CA19-9 glycans and the limitation of analytical methods. Therefore, it is hard to characterize the real CA19-9 carrier protein and distinguish their variation from either glycan or protein expression. This project aims to establish a highly sensitive method to comprehensively analyze the CA19-9 carrier glycoproteome. For identification and quantification of CA19-9 glycosylation, the method was developed via the establishment of integrated sample preparation to enrich CA19-9 peptides, optimization of LC-MS method and characterization of CA19-9 glycan and peptide structures. The CA19-9 glycosylation of pancreatic cancer was characterized by the developed method, and better markers were screened after comparison with other non-tumor tissues. If the project would be implemented smoothly, it is expected the multi-dimensional quantization of CA19-9 was greatly improved, and a new strategy would be provided for the early diagnosis of pancreatic cancer.