Differential Proteome Analysis of Extracellular Vesicles from Breast Cancer Cell Lines by Chaperone Affinity Enrichment.
Differential Proteome Analysis of Extracellular Vesicles from Breast Cancer Cell Lines by Chaperone Affinity Enrichment.
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DOI：
10.3390/proteomes5040025
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发表时间：
2017-10-08
期刊：
影响因子：
3.3
通讯作者：
Doucette AA
中科院分区：
文献类型：
--
作者：
Griffiths SG;Cormier MT;Clayton A;Doucette AA
人体组织液的复杂性妨碍了通过免疫测定或质谱法及时鉴定癌症生物标志物。越来越有吸引力的策略是主要富集与正常细胞相比以加速方式从癌细胞释放的细胞外囊泡（EV）。Vn 96肽在本文中用于从乳腺癌的细胞模型中回收释放到培养基中的EV的子集。Vn 96对装饰EV表面的热休克蛋白（HSP）具有亲和力。反映它们的起源细胞，癌症EV显示出与正常表型的离散差异。GELFrEE LC/MS从所有三种电动汽车来源中鉴定出了广泛的蛋白质组，其中绝大多数之前已在ExoCarta数据库中报道过。Vn 96亲和蛋白质组的途径分析明确区分了EV与致瘤细胞系（SKBR 3和MCF-7）相对于非致瘤来源（MCF-10a），特别是关于改变的代谢酶，信号传导和伴侣蛋白。蛋白质数据集提供了来自培养细胞脱落物质的有价值的信息。很可能使用本文所述的方法可以从建立的和原代细胞培养物收集大量的生物标志物身份。
The complexity of human tissue fluid precludes timely identification of cancer biomarkers by immunoassay or mass spectrometry. An increasingly attractive strategy is to primarily enrich extracellular vesicles (EVs) released from cancer cells in an accelerated manner compared to normal cells. The Vn96 peptide was herein employed to recover a subset of EVs released into the media from cellular models of breast cancer. Vn96 has affinity for heat shock proteins (HSPs) decorating the surface of EVs. Reflecting their cells of origin, cancer EVs displayed discrete differences from those of normal phenotype. GELFrEE LC/MS identified an extensive proteome from all three sources of EVs, the vast majority having been previously reported in the ExoCarta database. Pathway analysis of the Vn96-affinity proteome unequivocally distinguished EVs from tumorigenic cell lines (SKBR3 and MCF-7) relative to a non-tumorigenic source (MCF-10a), particularly with regard to altered metabolic enzymes, signaling, and chaperone proteins. The protein data sets provide valuable information from material shed by cultured cells. It is probable that a vast amount of biomarker identities may be collected from established and primary cell cultures using the approaches described here.