A virtual-droplet system for sensing MMP9 activity of single suspended and adhered cancer cells
A virtual-droplet system for sensing MMP9 activity of single suspended and adhered cancer cells
复制标题
用于感测单个悬浮和粘附癌细胞的 MMP9 活性的虚拟液滴系统
DOI：
10.1016/j.snb.2020.127749
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发表时间：
2020-04-01
影响因子：
8.4
通讯作者：
Liu, Maili
中科院分区：
文献类型：
--
作者：
Li, Ying;Zhang, Pengchao;Liu, Maili
癌细胞的MMP 9活性在肿瘤发展中起着至关重要的作用。肿瘤细胞在细胞-细胞和细胞-基质相互作用期间处于粘附分布。然而，这对于当前的平台（例如，典型的液滴系统）来测量粘附谱中由单细胞分泌的MMP 9，并回收感兴趣的细胞用于下游分析。本文报道了一种基于“虚拟液滴”的单细胞蛋白酶传感器（SCPS），它可以满足上述两个要求。SCPS可以在一分钟内形成648个虚拟液滴，作为一个细胞/液滴的轮廓，这提供了一个具有基底的隔离空间来支持细胞的粘附，并能够在两小时内对数百个单个粘附细胞进行MMP 9测量。除了癌细胞在MMP 9产生中的高细胞异质性之外，我们还发现粘附的MDA-MB-231细胞比悬浮细胞产生相对更多的MMP 9。此外，回收产生高MMP 9和低MMP 9的细胞，并对其进行mRNA测序。结果表明，一些与细胞迁移和侵袭相关的KEGG途径被富集，并且一些相关基因（例如，FYN、MAPK 3、RAF）在高MMP 9细胞中上调。该系统为单细胞蛋白酶活性的测定提供了一种很有前途的方法。
Cancer cells' MMP9 activity plays vital roles in tumor development. Tumor cells are in an adhesion profile during cell-cell and cell-matrix interaction. However, it is challenging for current platforms (e.g., typical droplet systems) to measure MMP9 secreted by single cells in an adhesion profile and retrieve the cells of interest for downstream analysis. Herein, we report a single-cell protease sensor (SCPS) based on 'virtual droplet' that can fulfill the above two requirements. SCPS can form 648 virtual droplets within one minute as a one-cell-perdroplet profile, which offers an isolated space with a substrate to support cells' adhesion and enables MMP9 measurement for hundreds of single adhered cells within two hours. In addition to high cellular heterogeneity of cancer cells in MMP9 production, we also found that adhered MDA-MB-231 cells produced relatively more MMP9 than that done by suspended cells. Furthermore, the cells producing high MMP9 and low MMP9 were retrieved and mRNA-sequence was performed on them. The results demonstrated that some KEGG pathways associated with cell migration and invasion were enriched and a few related genes (e.g., FYN, MAPK3, RAF) were upregulated in high-MMP9 cells. This SCPS system provides a promising strategy for measuring single-cell's protease activity.