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Cancers (Basel) 10, E40. Regulation of cancer stem cell metabolism by secreted frizzled-related protein 4 (sFRP4). 2018

Deshmukh, A., Arfuso, F., Newsholme, P. and Dharmarajan, A.

Notes: Cancer stem cells (CSCs; 10,000 cells/well in 96-well plates) were washed with PBS and 50µl of 1mM 2-Deoxy-d-Glucose (2DG) added. After 90 minutes, glucose uptake was measured using the Glucose Uptake-Glo™ Assay. CSCs were isolated from cell lines and plated at 10,000 cells/well in 96-well plates. After growing in defined medium supplemented with various amounts of glucose plus glutamine and growth factors for 2 days, cells were treated for 24 hours and 2µl samples removed and added to 98µl of PBS. Extracellular metabolites were assessed using the Glutamine/Glutamate-Glo™ Assay. Isolated CSCs were plated at 10,000 cells/well in 96-well plates, grown for 3 days, treated for 24 hours, medium removed and cells lysed. The lysates were treated to preserve NAD+ and NADH before mixing with the NAD/NADH-Glo™ Detection Reagent and luminescence measured. (4959)

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SLAS Discov 4, 366–377. Bioluminescent Assays for Glucose and Glutamine Metabolism: High-Throughput Screening for Changes in Extracellular and Intracellular Metabolites 2017

Leippe, D., Sobol, M., Vidugiris, G., Cali, J.J. and Vidugiriene, J.

Notes: The authors of this paper describe bioluminescent glucose, lactate, glutamine and glutamate assays suitable for high-throughput analysis of two major metabolic pathways in cancer cells: glycolysis and glutaminolysis. (4769)

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Oncogene 36(15), 2074-2084. Glutamine activates STAT3 to control cancer cell proliferation independently of glutamine metabolism. 2017

Cacace, A., Sboarina, M., Vazeille, T., and Sonveaux, P.

Notes: These authors report that glutamine promotes proliferation of cancer cells independently of its use as a metabolic fuel or as a precursor of glutathione, and that glutamine also activates transcription factors hypoxia-inducible factor-1, mammalian target of rapamycin and c-Myc, but these factors do not mediate the effects of glutamine on cancer cell proliferation. During the study, total RNA was isolated from cells using the Maxwell® RSC instrument,  ATP levels were measured using the CellTiter-Glo® Assay and intracellular glutamine and glutamate were measured using the Glutamine/Glutamate-Glo™ Assay. HIF-1 and PHD activities were measured using the Dual-Luciferase® Reporter Assay.


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