微泡
小RNA
液体活检
细胞外小泡
生物分子
癌症
分离(微生物学)
材料科学
胞外囊泡
癌症研究
计算生物学
诊断生物标志物
外体
纳米技术
生物
生物信息学
生物化学
细胞生物学
基因
遗传学
作者
Minjeong Jang,Giwoong Choi,Yoon Young Choi,Jae Eun Lee,Jik-Han Jung,Seungwon Oh,Dai Hoon Han,Haeshin Lee,Ji-Ho Park,Jae Ho Cheong,Pilnam Kim
标识
DOI:10.1038/s41427-019-0184-0
摘要
Abstract Small extracellular vesicles (EVs), including exosomes, in body fluids have important applications in the noninvasive liquid biopsy-based diagnosis of cancer. Current EV-based diagnostic techniques still face practical challenges, such as inefficient EV isolation. Here, we report an efficient, resource-free pre-enrichment approach using (–)-epigallocatechin-3-gallate (EGCG), a polyphenolic biomolecule, to isolate and detect exosomal microRNAs (miRNAs) in human blood plasma samples. Our system comprises three steps: (1) EGCG-mediated EV aggregation, (2) filter-based EV isolation, and (3) molecular beacon-based detection of target miRNA in EVs. Using blood samples from cancer patients with gastric cancer or hepatocellular carcinoma, we constructed an EGCG-assisted miRNA diagnostic system. For both cancers, the levels of target miRNAs (miR-21, -27a, and -375) in EVs were strongly correlated with those in the publicly available GEO database. Our approach, an easy-to-use method for efficient EV isolation and the detection of miRNA in clinical samples, is applicable for molecular diagnostics in precision medicine.
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