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

Abstract

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 (13) C6 -glucose and heteronuclear two-dimensional (2D) NMR. The method allowed for metabolomic differentiation between cancer and normal cells on the basis of time-dependent 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.

Recent Advances in NMR-Based Metabolomics

“…43 This method allowed for metabolic differentiation between cancer and normal cells on the basis of time-dependent changes in metabolite concentrations. The anticancer effects of galloflavin were also monitored, which revealed previously unknown functional targets of galloflavin.…”

Characterization of proteins by in-cell NMR spectroscopy in cultured mammalian cells

“…3-mm Shigemi tubes can be used in 5-mm probes with a proper spinner/adaptor. If more cells are available, larger tubes (5 mm normal/Shigemi tubes) may be used to increase the signalto-noise ratio (S/N) of the experiments, as shown elsewhere 34,35 .…”

Plasmonic silver nanoshells for drug and metabolite detection

“…I n-vitro metabolic diagnosis relies on designed materials-based analytical platforms for detection of selected metabolites in biological samples, which has a key role for disease detection and therapeutic evaluation in clinics 1,2 . Currently, both inorganic and organic materials have been used for metabolic analysis by selected spectroscopy methods, including nuclear magnetic resonance spectroscopy, biochemical analyzers, and mass spectrometry (MS) et al [1][2][3][4][5] . In contrast with other methods, MS enjoys unique advantages of high accuracy, sensitivity, resolution, and throughput.…”