Analysis of Glucose Uptake by Highly Metastatic Nanog-overexpressing Melanoma Cells Using Fluorescent Glucose Analogs

Fluorescent glucose analogs, 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-D-glucose (2-NBDG) and 2-[N-(7-nitrobenz-2-oxa-1,3- diazol-4-yl)amino]-2-deoxy-L-glucose (2-NBDLG) were applied to the detection of metastasis dependent glucose uptake and intracellular movement. The uptakes of both analogs by highly metastatic mouse melanoma cell line, Nanog+BL6 were significantly greater than those by the control cell line, BL6. Nucleus shape in Nanog+BL6, however, was not influenced by high concentration of 2-NBDLG, suggesting less active state of 2NBDLG in cells than in other cells reported elsewhere. We focused the involvement of Slc37a4 that was the top rank gene among those up -regulated by Nanog overexpression. When Slc37a4 in Nanog+BL6 was knocked down, the uptake of 2-NBDG decreased to 62% but 2-NBDLG uptake was unchanged. This suggests that combined use of 2-NBDG and 2-NBDLG can detect intracellular glucose movement relevant to cancer cells with different metastatic potentials.