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化学进展 2019, Vol. 31 Issue (6): 847-857 DOI: 10.7536/PC181027 前一篇   后一篇

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肿瘤外泌体的分析检测

乔斌, 陈虹妃, 张卉**(), 蔡称心   

  1. 江苏省新型动力电池重点实验室 南京师范大学化学与材料科学学院 南京 210023
  • 收稿日期:2018-10-23 出版日期:2019-06-15 发布日期:2019-04-26
  • 通讯作者: 张卉
  • 基金资助:
    国家自然科学基金项目(21375063); 江苏省自然科学基金面上项目(BK20181382)
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Analysis and Detection of Tumor Exosomes

Bin Qiao, Hongfei Chen, Hui Zhang**(), Chenxin Cai   

  1. Jiangsu Key Laboratory of New Power Batteries, College of Chemical and Materials Science, Nanjing Normal University, Nanjing 210023, China
  • Received:2018-10-23 Online:2019-06-15 Published:2019-04-26
  • Contact: Hui Zhang
  • About author:
    ** E-mail:
  • Supported by:
    National Natural Science Foundation of China(21375063); Natural Science Foundation of Jiangsu Province(BK20181382)

肿瘤外泌体是由肿瘤细胞释放到细胞外环境的微小囊泡,其直径约为30~150 nm,主要存在于血液、尿液、唾液等多种体液中,是早期肿瘤诊断的标志物之一。肿瘤外泌体具有较好的稳定性且含量丰富,因此是液体活检标志物的研究热点。肿瘤外泌体携带母细胞相关的蛋白、脂质和核酸等生物活性物质,为生物检测提供了多种特征标志物。本文就肿瘤外泌体的生成、分离、表征及分析检测进行了论述,重点讨论了肿瘤外泌体检测的研究进展。

关键词： 外泌体, 表面蛋白, 液体活检, 分离, 分析检测

Tumor exosomes are nanoscale membrane-enclosed vesicles with a diameter of 30~150 nm released from tumor cells into the extracellular environment. They are present in human blood, urine, saliva and many other body fluids and are one of the biomarkers for early tumor diagnosis. Tumor exosomes have the characteristics of good stability and abundance, which is a hot spot in liquid biopsy. Tumor exosomes carry a considerable amount of molecular information derived from the originating cell such as proteins, lipids and nucleic acids, providing a variety of characteristic biomarkers that can be used for cancer detection. In this paper, the formation, isolation, characterization and detection of tumor exosomes are discussed, and the research progress of the detection of tumor exosomes is discussed in detail.

Key words: exosomes, surface proteins, liquid biopsy, isolation, analysis and detection
()
图1 外泌体的结构[6]
Fig. 1 Structure of exosome[6]. Copyright 2018 American Chemical Society.
图2 外泌体的生成[6]
Fig. 2 Schematic illustration of exosome biogenesis[6]. Copyright 2018 American Chemical Society.
图3 各种电子显微镜的外泌体表征图。(a)扫描电子显微镜[47](b)透射电子显微镜[48](c)冷冻电子显微镜[50](d)原子力显微镜[51]
Fig. 3 Various micrographs of exosomes.(a) Scanning electron microscopy(SEM)[47]. Copyright 2012 Nature Publishing Group.(b) Transmission electron microscopy(TEM)[48]. Copyright 2018 American Chemical Society. (c) Cryo-electron microscopy(Cryo-EM)[50]. Copyright 2013 Taylor & Francis Ltd.(d) Atomic force microscopy(AFM)[51]. Copyright 2010 American Chemical Society.
图4 采用免疫亲和分离和基于胆固醇的信号放大相结合的方法对外泌体进行定量的原理图[78]
Fig. 4 Working principle of the proposed method for exosome quantifi cation by a combination of immunoaffinity separation and cholesterol-based signal amplification[78]. Copyright 2017 American Chemical Society.
图5 一种基于铜介导的信号放大的方法用于检测肿瘤细胞分泌的外泌体的工作原理图[83]
Fig. 5 Working principle of the proposed method for exosome detection based on a copper-mediated signal amplification strategy[83]. Copyright 2018 American Chemical Society.
图6 基于表面增强拉曼(SERS)检测肺癌细胞外泌体的原理图[86]。(a,b)肺癌细胞和正常细胞通过多囊泡内涵体与细胞质膜融合向细胞外环境释放外泌体。(c,d)肺癌细胞和正常细胞外泌体的拉曼光谱。(e)用面板c和d的方法获得了SERS光谱。红线代表着肺癌细胞外泌体的特定光谱峰。(f)外泌体的分类是通过SERS光谱的主成分分析(PCA)进行的
Fig. 6 Schematic diagram of lung cancer diagnosis by SERS classification of exosome[86].(a, b) Lung cancer cell and normal cell release exosomes to the extracellular environment having their own profiles by fusing multivesicular endosomes to plasma membrane, respectively.(c, d) Raman spectra of lung cancer cell and normal cell derived exosomes were achieved by SERS respectively.(e) SERS spectra achieved by methods of panels c and d are shown. Red lines indicate specific peaks of lung cancer derived exosomes.(f) Exosome classification is done by PCA of SERS spectra. Copyright 2017 American Chemical Society.
图7 外泌体的检测图[48]。(A) 适配体识别结合外泌体,释放mDNAs。(B)检测释放的mDNAs;(a)P1修饰金电极,并用MCH封闭;(b)捕捉mDNAs;(c)核酸外切酶Ⅲ作用;(d)DPV电化学检测
Fig. 7 Detection of tumor exosomes[48].(A) Aptamers recognize and combine with exosomes and release mDNAs.(B) Detection of the released mDNAs:(a) P1-modified gold electrode blocked with MCH,(b) capture of mDNAs,(c) treated with Exo Ⅲ exonuclease,(d) DPV measurement in the electrochemical buffer. Copyright 2018 American Chemical Society.
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摘要

肿瘤外泌体的分析检测