Identification and proteomic analysis of osteoblast-derived exosomes
Identification and proteomic analysis of osteoblast-derived exosomes
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成骨细胞来源的外泌体的鉴定和蛋白质组分析
DOI：
10.1016/j.bbrc.2015.09.135
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发表时间：
2015-11-06
影响因子：
3.1
通讯作者：
Mu, Xiongzheng
中科院分区：
文献类型：
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作者：
Ge, Min;Ke, Ronghu;Mu, Xiongzheng
外泌体是具有细胞间通讯功能的纳米大小的囊泡，由各种细胞类型释放。为了揭示成骨细胞外泌体的相关知识，探索其潜在的成骨功能，我们采用超离心方法从小鼠Mc3t3上清液中分离出微泡，通过电镜和免疫印迹技术对外泌体进行了表征，并通过蛋白质组学分析获得了外泌体的蛋白质谱。结果表明，微泡直径在30 ~ 100 nm之间，形状圆形，呈杯状凹，表达外泌体标记物肿瘤易感基因（TSG） 101和漂浮蛋白（fllot） 1。共鉴定出1069个蛋白，其中786个蛋白与ExoCarta数据库重叠。基因肿瘤学分析表明，外泌体主要来源于质膜，主要参与蛋白质定位和细胞内信号转导。独创性途径分析显示，途径主要涉及外泌体的生物发生、形成、摄取和成骨。其中真核起始因子2通路在成骨过程中起重要作用。我们的研究确定了成骨细胞来源的外泌体，揭示了它们的含量，提出了潜在的成骨相关蛋白和途径，并提供了丰富的蛋白质组学数据资源，这将为进一步研究单个蛋白在骨病中的功能提供宝贵的价值。(C) 2015爱思唯尔公司版权所有。
Exosomes are nanometer-sized vesicles with the function of intercellular communication, and they are released by various cell types. To reveal the knowledge about the exosomes from osteoblast, and explore the potential functions of osteogenesis, we isolated microvesicles from supernatants of mouse Mc3t3 by ultracentrifugation, characterized exosomes by electron microscopy and immunoblotting and presented the protein profile by proteomic analysis. The result demonstrated that microvesicles were between 30 and 100 nm in diameter, round shape with cup-like concavity and expressed exosomal marker tumor susceptibility gene (TSG) 101 and flotillin (Flot) 1. We identified a total number of 1069 proteins among which 786 proteins overlap with ExoCarta database. Gene Oncology analysis indicated that exosomes mostly derived from plasma membrane and mainly involved in protein localization and intracellular signaling. The Ingenuity Pathway Analysis showed pathways are mostly involved in exosome biogenesis, formation, uptake and osteogenesis. Among the pathways, eukaryotic initiation factor 2 pathways played an important role in osteogenesis. Our study identified osteoblast-derived exosomes, unveiled the content of them, presented potential osteogenesis-related proteins and pathways and provided a rich proteomics data resource that will be valuable for further studies of the functions of individual proteins in bone diseases. (C) 2015 Elsevier Inc. All rights reserved.