文献名:Quantitative proteomic analysis of small and large extracellular vesicles (EVs) reveals enrichment of adhesion proteins in small EVs(小型和大型胞外囊泡的定量蛋白质组学分析揭示小型胞外囊泡中粘附蛋白的富集)

期刊名:Journal of proteome research

发表时间:2019年1月

单位:

  1. 范德比尔特大学医学院
  2. 新墨西哥大学
  3. 莎拉坎农研究所

技术:iTRAQ蛋白质组定量技术

分享人: 柯酩

一、概述:(用精炼的语言描述文章的整体思路及结果)

文章采用iTRAQ联合LC-MS/MS技术,对肿瘤细胞DKs8的大型胞外囊泡(large extracellular vesicles,LEVs)和小型胞外囊泡(SEVs)的蛋白质组进行定量差异对比,揭示了SEVs,而不是LEVs,的一种主要功能是促进细胞-细胞和细胞-基质的粘附作用。

二、研究背景:(简要介绍研究进展动态、研究目的和意义)

SEVs的主要成分是外泌体(exosomes),而LEVs的主要成分是微囊泡(microvesicles,MVs),典型的外泌体直径在30到150纳米之间,而典型的微囊泡直径达到150到1000纳米。拯救实验(rescue experiment)表明,只有SEVs能拯救由于外泌体分泌抑制造成的细胞运动缺陷和HNSCC的致癌作用,影响肿瘤细胞的血清非依赖生长和侵润。这表明小型胞外囊泡和大型胞外囊泡装载不同的分子从而介导不同的功能。本文旨在对比LEVs和SEVs中蛋白质的定量差异,从而揭示它们功能差异的分子基础。

三、研究成果:(重点图表展示)

1、LEVs和SEVs的分离纯化以及蛋白质组学表征

A)Wetern blot检测DKs8全细胞、LEVs和密度梯度离心各组分中的标志蛋白:Flotillin,Hsp70,Tsg101和CD63。B)LEVs和SEVs的扫描电镜图。C)韦恩图展示三次iTRAQ标记定量检测到的蛋白质数目。D)火山图展示显著差异蛋白质(差异倍数>2,p<0.05)

2、Western blot验证所选蛋白在DKs8和HT1080EV中LEVs和SEVs的 3、SEVs促进SDs8的细胞粘附

3、SEVs促进SDs8的细胞粘附

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