Jianchen Yang1, Tessa Davis1, Anum S Kazerouni1, Yuan-I Chen1, Meghan J Bloom1, Hsin-Chih Yeh1,2, Thomas E Yankeelov1,3,4,5,6,7, John Virostko8,9,10. 1. Department of Biomedical Engineering, The University of Texas At Austin, Austin, TX, 78712, USA. 2. Texas Materials Institute, The University of Texas At Austin, Austin, TX, 78712, USA. 3. Department of Diagnostic Medicine, The University of Texas At Austin, 201 E. 24th Street, 1 University Station (C0200), Austin, TX, 78712, USA. 4. Department of Oncology, The University of Texas At Austin, Austin, TX, 78712, USA. 5. Oden Institute for Computational Engineering and Sciences, The University of Texas At Austin, Austin, TX, 78712, USA. 6. Livestrong Cancer Institutes, The University of Texas At Austin, Austin, TX, 78712, USA. 7. Department of Imaging Physics, The University of Texas MD Anderson Cancer Center, Houston, TX, 77030, USA. 8. Department of Diagnostic Medicine, The University of Texas At Austin, 201 E. 24th Street, 1 University Station (C0200), Austin, TX, 78712, USA. jack.virostko@austin.utexas.edu. 9. Department of Oncology, The University of Texas At Austin, Austin, TX, 78712, USA. jack.virostko@austin.utexas.edu. 10. Livestrong Cancer Institutes, The University of Texas At Austin, Austin, TX, 78712, USA. jack.virostko@austin.utexas.edu.
Abstract
PURPOSE: The reprogramming of cellular metabolism is a hallmark of cancer. The ability to noninvasively assay glucose and lactate concentrations in cancer cells would improve our understanding of the dynamic changes in metabolic activity accompanying tumor initiation, progression, and response to therapy. Unfortunately, common approaches for measuring these nutrient levels are invasive or interrupt cell growth. This study transfected FRET reporters quantifying glucose and lactate concentration into breast cancer cell lines to study nutrient dynamics and response to therapy. PROCEDURES: Two FRET reporters, one assaying glucose concentration and one assaying lactate concentration, were stably transfected into the MDA-MB-231 breast cancer cell line. Correlation between FRET measurements and ligand concentration were measured using a confocal microscope and a cell imaging plate reader. Longitudinal changes in glucose and lactate concentration were measured in response to treatment with CoCl2, cytochalasin B, and phloretin which, respectively, induce hypoxia, block glucose uptake, and block glucose and lactate transport. RESULTS: The FRET ratio from the glucose and lactate reporters increased with increasing concentration of the corresponding ligand (p < 0.005 and p < 0.05, respectively). The FRET ratio from both reporters was found to decrease over time for high initial concentrations of the ligand (p < 0.01). Significant differences in the FRET ratio corresponding to metabolic inhibition were found when cells were treated with glucose/lactate transporter inhibitors. CONCLUSIONS: FRET reporters can track intracellular glucose and lactate dynamics in cancer cells, providing insight into tumor metabolism and response to therapy over time.
PURPOSE: The reprogramming of cellular metabolism is a hallmark of cancer. The ability to noninvasively assay glucose and lactate concentrations in cancer cells would improve our understanding of the dynamic changes in metabolic activity accompanying tumor initiation, progression, and response to therapy. Unfortunately, common approaches for measuring these nutrient levels are invasive or interrupt cell growth. This study transfected FRET reporters quantifying glucose and lactate concentration into breast cancer cell lines to study nutrient dynamics and response to therapy. PROCEDURES: Two FRET reporters, one assaying glucose concentration and one assaying lactate concentration, were stably transfected into the MDA-MB-231 breast cancer cell line. Correlation between FRET measurements and ligand concentration were measured using a confocal microscope and a cell imaging plate reader. Longitudinal changes in glucose and lactate concentration were measured in response to treatment with CoCl2, cytochalasin B, and phloretin which, respectively, induce hypoxia, block glucose uptake, and block glucose and lactate transport. RESULTS: The FRET ratio from the glucose and lactate reporters increased with increasing concentration of the corresponding ligand (p < 0.005 and p < 0.05, respectively). The FRET ratio from both reporters was found to decrease over time for high initial concentrations of the ligand (p < 0.01). Significant differences in the FRET ratio corresponding to metabolic inhibition were found when cells were treated with glucose/lactate transporter inhibitors. CONCLUSIONS: FRET reporters can track intracellular glucose and lactate dynamics in cancer cells, providing insight into tumor metabolism and response to therapy over time.
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