癌细胞
葡萄糖氧化酶
厌氧糖酵解
糖酵解
细胞内
化学
生物化学
葡萄糖摄取
癌症
葡萄糖转运蛋白
碳水化合物代谢
细胞培养
生物物理学
纳米传感器
生物
新陈代谢
生物传感器
纳米技术
内分泌学
材料科学
胰岛素
遗传学
作者
Raphael Ap Sanches Nascimento,Rıfat Emrah Özel,Wai H. Mak,Marcelo Mulato,Bakthan Singaram,Nader Pourmand
出处
期刊:Nano Letters
[American Chemical Society]
日期:2016-01-19
卷期号:16 (2): 1194-1200
被引量:146
标识
DOI:10.1021/acs.nanolett.5b04495
摘要
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.
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