Protein Profiling and Sizing of Extracellular Vesicles from Colorectal Cancer Patients via Flow Cytometry
Protein Profiling and Sizing of Extracellular Vesicles from Colorectal Cancer Patients via Flow Cytometry
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通过流式细胞术对结直肠癌患者的细胞外囊泡进行蛋白质分析和大小测定
DOI：
10.1021/acsnano.7b07782
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发表时间：
2018-01-01
期刊：
影响因子：
17.1
通讯作者：
Yan, Xiaomei
中科院分区：
文献类型：
--
作者：
Tian, Ye;Ma, Ling;Yan, Xiaomei
细胞外囊泡（EV）已经激发了相当大的科学和临床兴趣，但是由于它们的小颗粒尺寸、低丰度的蛋白质和整体异质性，单个EV的蛋白质谱分析和大小测定仍然具有挑战性。在实验室构建的高灵敏度流式细胞仪（HSFCM）的基础上，我们在这里报告了一种快速的方法，用于定量分析单个EV的多参数分析，分析速率高达每分钟10 000个颗粒。在几分钟内获得了统计学上稳健的粒度分布，其分辨率和轮廓与cryo-TEM测量结果匹配良好。在免疫荧光染色后定量表达⑶ 9、⑶ 63和/或⑶ 81的EV亚群。当HSFCM用于分析血液样品时，与健康对照相比，在结直肠癌患者中鉴定出显著升高的CD 147阳性EV水平（P < 0.001）。HSFCM为单个EV的表面蛋白质谱分析和大小测定提供了一个灵敏和快速的平台，这可以极大地帮助理解EV介导的细胞间通讯以及先进的诊断和治疗策略的开发。
Extracellular vesicles (EVs) have stimulated considerable scientific and clinical interest, yet protein profiling and sizing of individual EVs remains challenging due to their small particle size, low abundance of proteins, and overall heterogeneity. Building upon a laboratory-built high-sensitivity flow cytometer (HSFCM), we report here a rapid approach for quantitative multiparameter analysis of single EVs down to 40 nm with an analysis rate up to 10 000 particles per minute. Statistically robust particle size distribution was acquired in minutes with a resolution and profile well matched with those of cryo-TEM measurements. Subpopulations of EVs expressing CD9, CD63, and/or CD81 were quantified upon immunofluorescent staining. When HSFCM was used to analyze blood samples, a significantly elevated level of CD147-positive EVs was identified in colorectal cancer patients compared to healthy controls (P < 0.001). HSFCM provides a sensitive and rapid platform for surface protein profiling and sizing of individual EVs, which could greatly aid the understanding of EV-mediated intercellular communication and the development of advanced diagnostic and therapeutic strategies.