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Molecular Imaging and Biology

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01-10-2011 | Research Article

Development of High-Throughput Quantitative Assays for Glucose Uptake in Cancer Cell Lines

Authors: Mohamed Hassanein, Brandy Weidow, Elizabeth Koehler, Naimish Bakane, Shawn Garbett, Yu Shyr, Vito Quaranta

Published in: Molecular Imaging and Biology | Issue 5/2011

Abstract

Purpose

Metabolism, and especially glucose uptake, is a key quantitative cell trait that is closely linked to cancer initiation and progression. Therefore, developing high-throughput assays for measuring glucose uptake in cancer cells would be enviable for simultaneous comparisons of multiple cell lines and microenvironmental conditions. This study was designed with two specific aims in mind: the first was to develop and validate a high-throughput screening method for quantitative assessment of glucose uptake in “normal” and tumor cells using the fluorescent 2-deoxyglucose analog 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxyglucose (2-NBDG), and the second was to develop an image-based, quantitative, single-cell assay for measuring glucose uptake using the same probe to dissect the full spectrum of metabolic variability within populations of tumor cells in vitro in higher resolution.

Procedure

The kinetics of population-based glucose uptake was evaluated for MCF10A mammary epithelial and CA1d breast cancer cell lines, using 2-NBDG and a fluorometric microplate reader. Glucose uptake for the same cell lines was also examined at the single-cell level using high-content automated microscopy coupled with semi-automated cell-cytometric image analysis approaches. Statistical treatments were also implemented to analyze intra-population variability.

Results

Our results demonstrate that the high-throughput fluorometric assay using 2-NBDG is a reliable method to assess population-level kinetics of glucose uptake in cell lines in vitro. Similarly, single-cell image-based assays and analyses of 2-NBDG fluorescence proved an effective and accurate means for assessing glucose uptake, which revealed that breast tumor cell lines display intra-population variability that is modulated by growth conditions.

Conclusions

These studies indicate that 2-NBDG can be used to aid in the high-throughput analysis of the influence of chemotherapeutics on glucose uptake in cancer cells.

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Title: Development of High-Throughput Quantitative Assays for Glucose Uptake in Cancer Cell Lines
Authors: Mohamed Hassanein
Brandy Weidow
Elizabeth Koehler
Naimish Bakane
Shawn Garbett
Yu Shyr
Vito Quaranta
Publication date: 01-10-2011
Publisher: Springer-Verlag
Published in: Molecular Imaging and Biology / Issue 5/2011
Print ISSN: 1536-1632
Electronic ISSN: 1860-2002
DOI: https://doi.org/10.1007/s11307-010-0399-5

At a glance: The STEP trials

Springer Medicine

Development of High-Throughput Quantitative Assays for Glucose Uptake in Cancer Cell Lines

Abstract

Purpose

Procedure

Results

Conclusions

At a glance: The STEP trials

Springer Medicine

Abstract

Purpose

Procedure

Results

Conclusions

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