Characterization and proteomic profiling of pancreatic cancer-derived serum exosomes
Characterization and proteomic profiling of pancreatic cancer-derived serum exosomes
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胰腺癌衍生血清外泌体的表征和蛋白质组分析
DOI：
10.1002/jcb.27465
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发表时间：
2019-01-01
影响因子：
4
通讯作者：
Xiao, Ming-B.
中科院分区：
文献类型：
--
作者：
Jiao, Yu-J.;Jin, Dan-D.;Xiao, Ming-B.
胰腺癌（PC）是世界上已知的最致命的癌症之一，并且其在大多数患者中的预后很差。外泌体是从各种细胞类型释放的纳米尺寸的细胞外囊泡。它们参与细胞通讯。外泌体的应用大大改善了PC的诊断和治疗。在这项研究中，我们分离了PC衍生的外泌体，并研究了它们的蛋白质组学谱。然后，我们对蛋白质组数据进行了生物信息学分析。进行差示超离心以从人血清样品和四种PC细胞系中分离外来体。使用透射电子显微镜和Western印迹分析来表征分离的外泌体。采用液相色谱-串联质谱法鉴定血清外泌体蛋白质组。蛋白质组学分析表明，所有血清外泌体均来源于三组人类受试者;这些血清外泌体共含有655种蛋白质，其中315种蛋白质与ExoCarta数据库重叠。基因肿瘤学和京都基因百科全书和基因组分析提供了蛋白质组的功能注释。有趣的是，与分别来自健康志愿者或胰腺炎患者的血清外泌体相比，来自PC患者的血清外泌体中有18或14种蛋白质上调，11或14种蛋白质下调。膜联蛋白A11是一种钙依赖性磷脂结合蛋白，分别在PC细胞系（CFPAC-1）来源的外泌体和PC患者的肿瘤组织中表达。我们的数据为进一步研究PC衍生的外泌体的蛋白质组成及其参与PC生物学提供了基础。
Pancreatic cancer (PC) is one of the most lethal cancers known worldwide, and its prognosis is poor in most patients. Exosomes are nanosized extracellular vesicles, which are released from various cell types. They are involved in cellular communication. The diagnosis and treatment of PC were improved substantially with exosomes. In this study, we isolated PC‐derived exosomes and investigated their proteomic profile. Then, we conducted bioinformatic analysis on proteomic data. Differential ultracentrifugation was performed to isolate exosomes from human serum samples and four PC cell lines. Transmission electron microscopy and Western blot analysis were used to characterize the isolated exosomes. Liquid chromatography coupled with tandem mass spectrometry was conducted to identify the proteome of serum exosomes. Proteomic analysis demonstrated that all the serum exosomes were derived from three cohorts of human subjects; these serum exosomes contained a total of 655 proteins, out of which 315 proteins overlapped with ExoCarta database. Gene oncology and kyoto encyclopedia of genes and genomes analyses provided the functional annotation of the proteome. Interestingly, 18 or 14 proteins were upregulated and 11 or 14 proteins were downregulated in serum exosomes derived from patients with PC as compared with in serum exosomes derived from healthy volunteers or from pancreatitis patients respectively. Annexin A11, a calcium‐dependent phospholipid‐binding protein, was expressed in a PC cell line (CFPAC‐1)‐derived exosomes and in tumor tissues of patients with PC, respectively. Our data provided a basic foundation for further studies on the protein composition of PC‐derived exosomes and its involvement in PC biology.