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采用液相色谱/串联质谱技术对人胰管内乳头状黏液性肿瘤囊液蛋白质组进行分析。

Proteome characterization of human pancreatic cyst fluid from intraductal papillary mucinous neoplasm by liquid chromatography/tandem mass spectrometry.

机构信息

Department of Biomedical Engineering, Seoul National University College of Medicine, 103 Daehak-ro, Seoul, Korea.

Biomedical Research Institute, Seoul National University Hospital, 101 Daehak-ro, Seoul, Korea.

出版信息

Rapid Commun Mass Spectrom. 2017 Oct 30;31(20):1761-1772. doi: 10.1002/rcm.7959.

Abstract

RATIONALE

In recent years, the molecular components of pancreatic cyst fluid have been used for diagnosis and prognosis. Because the protein markers that are currently used in clinical tests are unreliable, proteomic studies to find new protein markers are being conducted. However, such researches have been limited due to the complexity of pancreatic cyst fluid and the immaturity of proteomic techniques.

METHODS

To overcome these limitations and provide a pancreatic cyst proteome dataset, we examined cyst fluid proteome with tandem mass spectrometry. The proteomic analysis was performed using a Orbitrap-based mass spectrometer (Q-Exactive) coupled with a 50-cm-long nano-liquid chromatography column. Protein mutations were identified using mutation sequence database search.

RESULTS

A total of 5850 protein groups were identified from microliters of cyst fluid. Among those, 3934 protein groups were reported for the first time in pancreatic cyst fluid. Although high-abundance proteins were not depleted in the experiment, our dataset detected almost all pancreatic tumor markers such as mucin family members, S100 proteins, and CEA-related proteins. In addition, 590 protein mutation marker candidates were discovered.

CONCLUSIONS

We provide a comprehensive cyst proteome dataset that includes cystic cellular proteins and mutated proteins. Our findings would serve as a rich resource for further IPMN studies and clinical applications. The MS data have been deposited in the ProteomeXchange with identifier PXD005671 (http://proteomecentral.proteomexchange.org/dataset/PXD005671).

摘要

背景

近年来,胰腺囊液的分子成分已被用于诊断和预后。由于目前临床检测中使用的蛋白质标志物不可靠,因此正在进行蛋白质组学研究以寻找新的蛋白质标志物。但是,由于胰腺囊液的复杂性和蛋白质组学技术的不成熟,此类研究受到限制。

方法

为了克服这些限制并提供胰腺囊液蛋白质组数据集,我们使用串联质谱法检查了囊液蛋白质组。蛋白质组分析是使用基于 Orbitrap 的质谱仪(Q-Exactive)与 50 厘米长的纳流色谱柱联用进行的。使用突变序列数据库搜索鉴定蛋白质突变。

结果

从微升的囊液中鉴定出 5850 个蛋白质组。其中,3934 个蛋白质组是首次在胰腺囊液中报道的。尽管实验中未耗尽高丰度蛋白质,但我们的数据集检测到了几乎所有胰腺肿瘤标志物,如粘蛋白家族成员、S100 蛋白和 CEA 相关蛋白。此外,还发现了 590 个蛋白质突变标志物候选物。

结论

我们提供了一个全面的囊液蛋白质组数据集,其中包括囊液细胞内蛋白质和突变蛋白质。我们的发现将为进一步的 IPMN 研究和临床应用提供丰富的资源。MS 数据已存入 ProteomeXchange,标识符为 PXD005671(http://proteomecentral.proteomexchange.org/dataset/PXD005671)。

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