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基于单细胞糖酵解的癌细胞亚群分离用微流控平台。

Microfluidic Platform for the Isolation of Cancer-Cell Subpopulations Based on Single-Cell Glycolysis.

机构信息

Department of Chemistry and Biochemistry, Santa Clara University, Santa Clara, California 95053, United States.

Department of Biology, Santa Clara University, Santa Clara, California 95053, United States.

出版信息

Anal Chem. 2020 May 19;92(10):6949-6957. doi: 10.1021/acs.analchem.9b05738. Epub 2020 Apr 30.

Abstract

High rates of glycolysis in tumors have been associated with cancer metastasis, tumor recurrence, and poor outcomes. In this light, single cells that exhibit high glycolysis are specific targets for therapy. However, the study of these cells requires efficient tools for their isolation. We use a droplet microfluidic technique developed in our lab, orting by nteracial ension (SIFT), to isolate cancer cell subpopulations based on glycolysis without the use of labels or active sorting components. By controlling the flow conditions on chip, the threshold of selection can be modified, enabling the isolation of cells with different levels of glycolysis. Hypoxia in tumors, that can be simulated with treatment with CoCl, leads to an increase in glycolysis, and more dangerous tumors. The device was used to enrich CoCl treated MDA-MB 231 breast cancer cells from an untreated population. It is also used to sort K562 human chronic myelogenous leukemia cells that have either been treated or untreated with 2-deoxy-d-glucose (2DG), a pharmaceutical that targets cell metabolism. The technique provides a facile and robust way of separating cells based on elevated glycolytic activity; a biomarker associated with cancer cell malignancy.

摘要

肿瘤中糖酵解率高与癌症转移、肿瘤复发和预后不良有关。从这个角度来看,表现出高糖酵解的单细胞是治疗的特定目标。然而,这些细胞的研究需要有效的分离工具。我们使用实验室开发的液滴微流控技术,通过界面张力(SIFT),在不使用标记物或主动分选组件的情况下,根据糖酵解分离癌细胞亚群。通过控制芯片上的流动条件,可以修改选择的阈值,从而实现分离具有不同糖酵解水平的细胞。肿瘤中的缺氧可以通过用 CoCl 处理来模拟,这会导致糖酵解增加,从而导致更危险的肿瘤。该设备用于从未处理的群体中富集用 CoCl 处理的 MDA-MB 231 乳腺癌细胞。它还用于分选 K562 人慢性髓性白血病细胞,这些细胞要么用 2-脱氧-D-葡萄糖(2DG)处理,要么未处理,2DG 是一种靶向细胞代谢的药物。该技术提供了一种简单而强大的基于升高的糖酵解活性分离细胞的方法;这是与癌细胞恶性程度相关的生物标志物。

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本文引用的文献

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Single-Cell Analysis Using Droplet Microfluidics.基于液滴微流控的单细胞分析。
Adv Biosyst. 2020 Jan;4(1):e1900188. doi: 10.1002/adbi.201900188. Epub 2019 Nov 26.
5
The Glycolytic Switch in Tumors: How Many Players Are Involved?肿瘤中的糖酵解转换:涉及多少参与者?
J Cancer. 2017 Sep 20;8(17):3430-3440. doi: 10.7150/jca.21125. eCollection 2017.
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Tumour acidosis: from the passenger to the driver's seat.肿瘤酸中毒:从乘客到驾驶座。
Nat Rev Cancer. 2017 Oct;17(10):577-593. doi: 10.1038/nrc.2017.77. Epub 2017 Sep 15.
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Cancer metabolism in space and time: Beyond the Warburg effect.癌症在时空上的代谢:超越沃伯格效应。
Biochim Biophys Acta Bioenerg. 2017 Aug;1858(8):556-572. doi: 10.1016/j.bbabio.2017.02.001. Epub 2017 Feb 4.

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