In Situ Simultaneous Detection of Surface Protein and microRNA in Clustered Extracellular Vesicles from Cancer Cell Lines Using Flow Cytometry

被引：3

作者：

Lee, Ji Yoon ^{[1
]}

Lee, Soye ^{[1
]}

Lee, Haneul ^{[1
]}

Tran, Tham Truong Phuong ^{[1
,2
]}

Kim, Byoung Choul ^{[1
,2
]}

Rhee, Won Jong ^{[1
,3
,4
]}

机构：

[1] Incheon Natl Univ, Dept Bioengn & Nanobioengn, Incheon 22012, South Korea

[2] Incheon Natl Univ, Dept Nanobioengineering, Incheon 22012, South Korea

[3] Incheon Natl Univ, Div Bioengn, Incheon 22012, South Korea

[4] Incheon Natl Univ, Res Ctr Bio Mat & Proc Dev, Incheon 22012, South Korea

来源：

ACS BIOMATERIALS SCIENCE & ENGINEERING | 2023年 / 9卷 / 11期

基金：

新加坡国家研究基金会;

关键词：

extracellular vesicles; cancer liquid biopsy; in situ simultaneous detection; flow cytometry; EV cluster; DELIVERY; EXOSOMES; SYSTEM; SIZE;

D O I：

10.1021/acsbiomaterials.3c01459

中图分类号：

TB3 [工程材料学]; R318.08 [生物材料学];

学科分类号：

0805 ; 080501 ; 080502 ;

摘要：

Extracellular vesicles (EVs) are becoming increasingly important in liquid biopsy for cancer because they contain multiple biomarkers, including proteins and RNAs, and circulate throughout the body. Cancer cell-derived EVs are highly heterogeneous, and multiplexed biomarker detection techniques are required to improve the accuracy of diagnosis. In addition, in situ EV biomarker detection increases the efficiency of the detection process because EVs are difficult to handle. In this study, in situ simultaneous detection of EV surface proteins, programmed cell death-ligand 1 (PD-L1), and internal miRNA-21 (miR-21) analyzed by conventional flow cytometry was developed for a breast cancer liquid biopsy. However, the majority of EVs were not recognized by flow cytometry for biomarker detection because the size of EVs was below the detectable size range of the flow cytometer. To solve this problem, the formation of EV clusters was induced by 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE)-polyethylene glycol-DSPE during biomarker detection. Consequently, both PD-L1 and miR-21 detection signals from cancer cell-derived EVs were drastically increased, making them distinguishable from normal cell-derived EVs. The in situ simultaneous cancer biomarker detection from EV clusters analyzed by flow cytometry contributes to an increase in the sensitivity and accuracy of the EV-based liquid biopsy for cancer.

引用

页码：6369 / 6378

页数：10

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