Proteome profiling of exosomes derived from human primary and metastatic colorectal cancer cells reveal differential expression of key metastatic factors and signal transduction components
Proteome profiling of exosomes derived from human primary and metastatic colorectal cancer cells reveal differential expression of key metastatic factors and signal transduction components
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DOI：
10.1002/pmic.201200562
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发表时间：
2013-05-01
期刊：
影响因子：
3.4
通讯作者：
Simpson, Richard J.
中科院分区：
文献类型：
--
作者：
Ji, Hong;Greening, David W.;Simpson, Richard J.
Exosome是细胞外直径40-100 nm的小膜囊，起源于晚期内噬菌体，可以介导RNA和蛋白质在细胞间的转移，以帮助转移前的生态位形成。使用原代(SW480)和转移性(SW620)人类等基因结直肠癌细胞系，我们比较了Exosome蛋白质谱，以获得对肿瘤进展基础的转移因素和信号分子的有价值的见解。OptiPrep密度梯度分级纯化的外切体直径为40-100 nm，浮力密度约为1.09g/mL，并显示了刻板的外切体标记TSG101、Alix和CD63。一个主要的发现是转移因子(MET、S100A8、S100A9、TNC)、信号转导分子(EFNB2、JAG1、SRC、TNIK)以及脂筏和脂筏相关成分(CAV1、FLOT1、FLOT2、PROM1)在转移性SW620细胞的外体中选择性地浓缩。此外，利用冷冻电子显微镜对外切体的超微结构成分进行了鉴定。本研究的一个关键发现是在结直肠癌细胞外体中检测到并共定位了蛋白复合体EpCAM-CLDN7和TNiK-RAP2A。转移结肠癌细胞来源的外切体中转移因子和信号通路成分的选择性浓缩有助于我们理解肿瘤与间质细胞在肿瘤微环境中的相互作用。
Exosomes are small extracellular 40-100 nm diameter membrane vesicles of late endosomal origin that can mediate intercellular transfer of RNAs and proteins to assist premetastatic niche formation. Using primary (SW480) and metastatic (SW620) human isogenic colorectal cancer cell lines we compared exosome protein profiles to yield valuable insights into metastatic factors and signaling molecules fundamental to tumor progression. Exosomes purified using OptiPrep density gradient fractionation were 40-100 nm in diameter, were of a buoyant density approximate to 1.09 g/mL, and displayed stereotypic exosomal markers TSG101, Alix, and CD63. A major finding was the selective enrichment of metastatic factors (MET, S100A8, S100A9, TNC), signal transduction molecules (EFNB2, JAG1, SRC, TNIK), and lipid raft and lipid raft-associated components (CAV1, FLOT1, FLOT2, PROM1) in exosomes derived from metastatic SW620 cells. Additionally, using cryo-electron microscopy, ultrastructural components in exosomes were identified. A key finding of this study was the detection and colocalization of protein complexes EPCAM-CLDN7 and TNIK-RAP2A in colorectal cancer cell exosomes. The selective enrichment of metastatic factors and signaling pathway components in metastatic colon cancer cell-derived exosomes contributes to our understanding of the cross-talk between tumor and stromal cells in the tumor microenvironment.