title: Development of high-throughput quantitative assays for glucose uptake in cancer cell lines.
abstract: OBJECTIVE
  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.
  
  
  METHODS
  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.affiliation: Vanderbilt Integrative Cancer Biology Center, Vanderbilt University Medical Center, Nashville, TN 37232, USA.
authors.forename: Mohamed
authors.initials: M
authors.lastname: Hassanein
authors.forename: Brandy
authors.initials: B
authors.lastname: Weidow
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Koehler
authors.forename: Naimish
authors.initials: N
authors.lastname: Bakane
authors.forename: Shawn
authors.initials: S
authors.lastname: Garbett
authors.forename: Yu
authors.initials: Y
authors.lastname: Shyr
authors.forename: Vito
authors.initials: V
authors.lastname: Quaranta
mesh_terms: D045744:Cell Line, Tumor; D005453:Fluorescence; D005947:Glucose / Q000378:metabolism*; D006801:Humans; D009369:Neoplasms / Q000378:metabolism*
chemical_list: D005947:Glucose
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