Real-Time Monitoring of Cancer Cell Metabolism and Effects of an Anticancer Agent using 2D In-Cell NMR Spectroscopy

Altered metabolism is a critical part of cancer cell properties, but real-time monitoring of metabolomic profiles has been hampered by the lack of a facile method. Here, we propose real-time metabolomic monitoring of live cancer cells using C-13(6)-glucose and heteronuclear two-dimensional (2D) NMR. The method allowed for metabolomic differentiation between cancer and normal cells on the basis of timedependent changes in metabolite concentrations. Cancer cells were found to have large in-and out-flux of pyruvate as well as increased net production of alanine and acetate. The method also enabled evaluation of the metabolic effects of galloflavin whose anticancer effects have been attributed to its specific inhibition of lactate dehydrogenase. Our approach revealed previously unknown functional targets of galloflavin, which were further confirmed at the protein levels. Our method is readily applicable to the study of metabolic alterations in other cellular disease model systems.