Facile preparation of salivary extracellular vesicles for cancer proteomics.
Facile preparation of salivary extracellular vesicles for cancer proteomics.
复制标题
轻松制备唾液细胞外囊泡用于癌症蛋白质组学
DOI：
10.1038/srep24669
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发表时间：
2016-04-19
影响因子：
4.6
通讯作者：
Xiao H
中科院分区：
文献类型：
--
作者：
Sun Y;Xia Z;Shang Z;Sun K;Niu X;Qian L;Fan LY;Cao CX;Xiao H
细胞外囊泡（EV）是细胞释放的膜包围结构，越来越多地被认为是细胞间通讯的介质。最近的报告表明，EV 参与重要的生物过程，并可以作为癌症生物标志物的潜在来源。作为一种有吸引力的 EV 来源，具有非侵入性的优点，人类唾液是临床诊断的独特介质。因此，我们提出了一种制备唾液细胞外囊泡（SEV）的简便方法。开发了结合过滤系统（ACCF）的亲和层析柱，可有效去除唾液中的高丰度蛋白质和粘性干扰物。通过该策略获得的 SEV 中的蛋白质分析与传统的离心方法进行了比较，结果表明可以多揭示约 70% 的 SEV 蛋白质。为了探索其在癌症蛋白质组学中的实用性，我们分析了肺癌患者和正常对照中 SEV 的蛋白质组。鸟枪法蛋白质组学分析表明，在癌症组和对照组中分别鉴定了 113 种和 95 种蛋白质。在这 63 种一直只在癌症组中发现的蛋白质中，有 12 种蛋白质与肺癌相关。我们的结果表明，通过所开发的策略制备的 SEV 是蛋白质组学的有价值的样本，并且可以作为癌症的有前途的液体活检。
Extracellular vesicles (EVs) are membrane surrounded structures released by cells, which have been increasingly recognized as mediators of intercellular communication. Recent reports indicate that EVs participate in important biological processes and could serve as potential source for cancer biomarkers. As an attractive EVs source with merit of non-invasiveness, human saliva is a unique medium for clinical diagnostics. Thus, we proposed a facile approach to prepare salivary extracellular vesicles (SEVs). Affinity chromatography column combined with filter system (ACCF) was developed to efficiently remove the high abundant proteins and viscous interferences of saliva. Protein profiling in the SEVs obtained by this strategy was compared with conventional centrifugation method, which demonstrated that about 70% more SEVs proteins could be revealed. To explore its utility for cancer proteomics, we analyzed the proteome of SEVs in lung cancer patients and normal controls. Shotgun proteomic analysis illustrated that 113 and 95 proteins have been identified in cancer group and control group, respectively. Among those 63 proteins that have been consistently discovered only in cancer group, 12 proteins are lung cancer related. Our results demonstrated that SEVs prepared through the developed strategy are valuable samples for proteomics and could serve as a promising liquid biopsy for cancer.