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从血浆中快速高效地分离和检测外泌体用于肺癌诊断。

Rapid and efficient isolation and detection of extracellular vesicles from plasma for lung cancer diagnosis.

机构信息

School of Medicine, Tsinghua University, Beijing 100084, China.

出版信息

Lab Chip. 2019 Jan 29;19(3):432-443. doi: 10.1039/c8lc01193a.

Abstract

Extracellular vesicles (EVs) are cell-derived nanoscale vesicles that provide promising biomarkers for the non-invasive diagnosis of cancer because they carry important cancer-related DNA, RNA and protein biomarkers. However, the clinical application of EVs is limited by tedious and non-standardized isolation methods that require bulky instrumentation. Here, we propose an easy-to-operate, simple dielectrophoretic (DEP) method for EV isolation with higher recovery efficiency (>83%) and higher purity than ultracentrifugation (UC). The DEP chip reduces the isolation procedure from 8 h to 30 min. To facilitate subsequent analysis, our DEP chip achieved integration of EV isolation and in situ lysis of EVs for the first time. Our chip also achieved on-chip siRNA delivery to EVs isolated by DEP. We found that EVs isolated from the plasma of lung cancer patients contained higher levels of miR-21, miR-191 and miR-192 compared to those from healthy people. With on-chip detection, EGFR in EVs could distinguish lung cancer patients from healthy people. Overall, this study provides an efficient and practical approach to the isolation and detection of EVs, which could be used for the early diagnosis of lung cancer.

摘要

细胞外囊泡(EVs)是源自细胞的纳米级囊泡,它们携带重要的与癌症相关的 DNA、RNA 和蛋白质生物标志物,为癌症的非侵入性诊断提供了有前景的生物标志物。然而,EVs 的临床应用受到繁琐和非标准化的分离方法的限制,这些方法需要庞大的仪器。在这里,我们提出了一种易于操作的简单介电泳(DEP)方法,用于 EV 分离,与超速离心(UC)相比,其回收率更高(>83%),纯度更高。DEP 芯片将分离过程从 8 小时缩短至 30 分钟。为了便于后续分析,我们的 DEP 芯片首次实现了 EV 分离和 EV 原位裂解的集成。我们的芯片还实现了在芯片上对通过 DEP 分离的 EV 进行 siRNA 递送。我们发现,与健康人相比,肺癌患者血浆中分离的 EV 含有更高水平的 miR-21、miR-191 和 miR-192。通过在芯片上检测,EV 中的 EGFR 可以区分肺癌患者和健康人。总体而言,这项研究提供了一种高效实用的 EV 分离和检测方法,可用于肺癌的早期诊断。

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