Proteomic Analysis of Exosomes and Exosome-Free Conditioned Media From Human Osteosarcoma Cell Lines Reveals Secretion of Proteins Related to Tumor Progression
Proteomic Analysis of Exosomes and Exosome-Free Conditioned Media From Human Osteosarcoma Cell Lines Reveals Secretion of Proteins Related to Tumor Progression
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DOI：
10.1002/jcb.25642
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发表时间：
2017-02-01
影响因子：
4
通讯作者：
Galindo, Mario
中科院分区：
文献类型：
--
作者：
Jerez, Sofia;Araya, Hector;Galindo, Mario
骨肉瘤是儿科患者中最常见的骨肿瘤，但也可能发生在以后的生活中。骨肿瘤有可能转移到肺，偶尔也会转移到其他重要器官。为了了解骨肉瘤细胞如何与其微环境相互作用以支持骨肿瘤的进展和转移，我们分析了三种人类骨肉瘤细胞系的分泌蛋白和外泌体。通过透射电镜（TEM）和免疫印迹检测特异性生物标志物（CD63、CD9和CD81），验证了外泌体的分离。外泌体蛋白和可溶性蛋白（小于100 kDa）通过nanoLC-MS/MS质谱分析鉴定，并通过功能基因本体聚类进行分类。我们为这两个部分鉴定了一组分泌组蛋白，并检测了三种骨肉瘤细胞系中常见或独特的蛋白。外泌体和游离外泌体蛋白质组的蛋白质本体比较揭示了与不同生物过程相关的功能类别的富集差异，包括与肿瘤进展相关的功能类别（即血管生成、细胞粘附和细胞迁移）。骨肉瘤细胞的分泌组特征与具有转移潜力的肿瘤细胞的病理特性一致。(C) 2016 Wiley期刊公司
Osteosarcomas are the most prevalent bone tumors in pediatric patients, but can also occur later in life. Bone tumors have the potential to metastasize to lung and occasionally other vital organs. To understand how osteosarcoma cells interact with their micro-environment to support bone tumor progression and metastasis, we analyzed secreted proteins and exosomes from three human osteosarcoma cell lines. Exosome isolation was validated by transmission electron microscopy (TEM) and immuno-blotting for characteristic biomarkers (CD63, CD9, and CD81). Exosomal and soluble proteins (less than 100 kDa) were identified by mass spectrometry analysis using nanoLC-MS/MS and classified by functional gene ontology clustering. We identified a secretome set of > 3,000 proteins for both fractions, and detected proteins that are either common or unique among the three osteosarcoma cell lines. Protein ontology comparison of proteomes from exosomes and exosome-free fractions revealed differences in the enrichment of functional categories associated with different biological processes, including those related to tumor progression (i.e., angiogenesis, cell adhesion, and cell migration). The secretome characteristics of osteosarcoma cells are consistent with the pathological properties of tumor cells with metastatic potential. (C) 2016 Wiley Periodicals, Inc.