A Bioluminescent Assay For Measuring Glucose Uptake

Glucose management is a key biological process. In adipose tissue and skeletal muscle, decreased rates of glucose uptake commonly associated with diabetes mellitus lead to chronic hyperglycemia and a wide array of physiological damage. In contrast, the overexpression of glucose transporters in cancer cells results in increased rates of glucose uptake to support their high rates of proliferation. Thus, activators and inhibitors of glucose uptake are useful for both diabetes management and anticancer therapies. The standard method of assaying glucose uptake involves addition of a radioactive glucose analog (2-deoxyglucose) and measurement of the accumulation of the stable and impermeable phosphorylated derivative, 2-deoxyglucose-6-phosphate (2DG6P). However, radioactive assays are laborious, costly, and require special handling of radioactive materials and waste. A simpler assay can be made by measuring the production of NADPH through the oxidation of 2DG6P by glucose-6-phosphate dehydrogenase. We have developed a bioluminescent glucose uptake assay that is both rapid and convenient and exhibits a larger signal window than comparable fluorescent or colorimetric approaches. The utility of this assay will be demonstrated with both cancer cells and insulin-sensitive 3T3L1 adipocytes and L6 myotubes. Citation Format: Michael P. Valley, Mary Sobol, Jolanta Vidugiriene, James J. Cali. A bioluminescent assay for measuring glucose uptake. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5435. doi:10.1158/1538-7445.AM2015-5435