Plasma extracellular vesicles detected by Single Molecule array technology as a liquid biopsy for colorectal cancer
Plasma extracellular vesicles detected by Single Molecule array technology as a liquid biopsy for colorectal cancer
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单分子阵列技术检测血浆细胞外囊泡作为结直肠癌液体活检
DOI：
10.1080/20013078.2020.1809765
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发表时间：
2020-01-01
影响因子：
16
通讯作者：
Li, Dawei
中科院分区：
文献类型：
--
作者：
Wei, Ping;Wu, Fei;Li, Dawei
循环细胞外囊泡（EVs）被认为是一种很有前途的诊断生物标志物。然而，在这一领域发表的研究有限，部分原因是能够检测细胞外囊泡的检测平台数量有限。本研究利用单分子阵列技术（SiMoa）建立了细胞外囊泡免疫检测方法，并对其在癌症诊断中的临床应用进行了评价。设计了两种细胞外囊泡检测方法CD9-CD63和Epcam-CD63，分别检测通用细胞外囊泡和肿瘤来源的细胞外囊泡。我们的研究结果表明，CD9-CD63和Epcam-CD63 SiMoa检测特异性检测细胞外囊泡，但不能检测高灵敏度的游离蛋白。在癌细胞培养液中检测到的Epcam-CD63水平与从同一癌细胞系中分离的表达epcam的ev水平一致，并通过Western blot检测。此外，化验区分癌和非癌血浆样品。与健康和良性对照相比，结直肠癌患者的CD9-CD63和Epcam-CD63信号最高。两种检测方法对结直肠癌的诊断均有较好的效果。此外，我们的研究结果表明，CD9-CD63检测是无进展生存期和总生存期的独立预后因素，而Epcam-CD63检测是OS的独立预后因素。
ABSTRACT Circulating extracellular vesicles (EVs) were recognized as a promising source of diagnostic biomarker. However, there are limited studies published in this area, partly due to the limited number of detection platforms capable of detecting extracellular vesicles. In this study, extracellular vesicle immunoassays were developed using the Single Molecule array technology (SiMoa) and their clinical applications to cancer diagnosis were evaluated. Two extracellular vesicle detection assays, CD9-CD63 and Epcam-CD63, were designed to detect universal extracellular vesicles and tumour-derived extracellular vesicles, respectively. Our results show that CD9-CD63 and Epcam-CD63 SiMoa assays specifically detect extracellular vesicles but not free proteins with high sensitivities. The Epcam-CD63 levels detected in cancer cell culture media were consistent with levels of Epcam-expressing EVs isolated from the same cancer cell lines and detected by Western blot. Furthermore, the assays distinguish cancerous from non-cancerous plasma samples. The highest CD9-CD63 and Epcam-CD63 signals were observed in colorectal cancer patients comparing to healthy and benign controls. Both assays showed superior diagnostic performance for colorectal cancer. In addition, our results show that CD9-CD63 detection is an independent prognosis factor for both progression free survival and overall survival, while Epcam-CD63 detectionis an independent prognosis factor for OS.