Document Detail


Development of high-throughput quantitative assays for glucose uptake in cancer cell lines.
MedLine Citation:
PMID:  20809209     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Mohamed Hassanein; Brandy Weidow; Elizabeth Koehler; Naimish Bakane; Shawn Garbett; Yu Shyr; Vito Quaranta
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural    
Journal Detail:
Title:  Molecular imaging and biology : MIB : the official publication of the Academy of Molecular Imaging     Volume:  13     ISSN:  1860-2002     ISO Abbreviation:  Mol Imaging Biol     Publication Date:  2011 Oct 
Date Detail:
Created Date:  2011-09-28     Completed Date:  2012-02-01     Revised Date:  2013-05-28    
Medline Journal Info:
Nlm Unique ID:  101125610     Medline TA:  Mol Imaging Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  840-52     Citation Subset:  IM    
Affiliation:
Vanderbilt Integrative Cancer Biology Center, Vanderbilt University Medical Center, Nashville, TN 37232, USA.
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MeSH Terms
Descriptor/Qualifier:
Cell Line, Tumor
Fluorescence
Glucose / metabolism*
Humans
Neoplasms / metabolism*
Grant Support
ID/Acronym/Agency:
U54 CA113007/CA/NCI NIH HHS; U54-CA113007/CA/NCI NIH HHS
Chemical
Reg. No./Substance:
50-99-7/Glucose
Comments/Corrections

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