Single Cell "Glucose Nanosensor" Verifies Elevated Glucose Levels in Individual Cancer Cells.
Single Cell "Glucose Nanosensor" Verifies Elevated Glucose Levels in Individual Cancer Cells.
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DOI：
10.1021/acs.nanolett.5b04495
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发表时间：
2016-02-10
期刊：
影响因子：
10.8
通讯作者：
Pourmand N
中科院分区：
文献类型：
--
作者：
Nascimento RA;Özel RE;Mak WH;Mulato M;Singaram B;Pourmand N
因为从氧化磷酸化到无氧糖酵解代谢的转变是癌症进展的标志，所以通过其独特的葡萄糖代谢特征来识别单个活癌细胞的方法将是有用的。在这里，我们提出了专门开发的纳米移液管，用于以时间和空间分辨率测量单细胞中的葡萄糖水平，我们使用这种技术来验证单个癌细胞确实可以显示更高的细胞内葡萄糖水平的假设。通过将葡萄糖氧化酶（GOx）共价固定到尖端，使葡萄糖与GOx的相互作用导致β-D-葡萄糖催化氧化为D-葡萄糖酸，将纳米移液管功能化为葡萄糖纳米传感器，这被测量为由于纳米移液管尖端处的介质的pH下降而引起的阻抗变化。校准研究表明，在尖端和溶液中的葡萄糖浓度的阻抗变化之间的直接关系。葡萄糖纳米传感器定量了人成纤维细胞和转移性乳腺癌细胞系MDA-MB-231和MCF 7中的单细胞细胞内葡萄糖水平，并显示与非恶性细胞相比，癌细胞表达了可重复和可靠的葡萄糖水平增加。纳米移液管允许对同一细胞进行重复采样，因为细胞在测量期间和测量后保持活力。因此，基于纳米移液管的葡萄糖传感器提供了一种将葡萄糖水平的变化与增殖或转移状态的变化进行比较的方法。该平台具有很大的潜力，机制的调查，作为一种诊断工具，以区分癌细胞从非恶性细胞在异质组织活检，以及用于监测癌症进展原位的工具。
Because the transition from oxidative phosphorylation to anaerobic glycolytic metabolism is a hallmark of cancer progression, approaches to identify single living cancer cells by their unique glucose metabolic signature would be useful. Here, we present nanopipettes specifically developed to measure glucose levels in single cells with temporal and spatial resolution, and we use this technology to verify the hypothesis that individual cancer cells can indeed display higher intracellular glucose levels. The nanopipettes were functionalized as glucose nanosensors by immobilizing glucose oxidase (GOx) covalently to the tip so that the interaction of glucose with GOx resulted in a catalytic oxidation of β-D-glucose to D-gluconic acid, which was measured as a change in impedance due to drop in pH of the medium at the nanopipette tip. Calibration studies showed a direct relationship between impedance changes at the tip and glucose concentration in solution. The glucose nanosensor quantified single cell intracellular glucose levels in human fibroblasts and the metastatic breast cancer lines MDA-MB-231 and MCF7 and revealed that the cancer cells expressed reproducible and reliable increases in glucose levels compared to the nonmalignant cells. Nanopipettes allow repeated sampling of the same cell, as cells remain viable during and after measurements. Therefore, nanopipette-based glucose sensors provide an approach to compare changes in glucose levels with changes in proliferative or metastatic state. The platform has great promise for mechanistic investigations, as a diagnostic tool to distinguish cancer cells from nonmalignant cells in heterogeneous tissue biopsies, as well as a tool for monitoring cancer progression in situ.