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 (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.
College of Pharmacy, Natural Product Research Institute, Seoul National University, Sillim-dong, Gwanak-gu, Seoul, 151-742 (Korea).
He
H
Wen
Yong Jin
YJ
An
Wen Jun
WJ
Xu
Keon Wook
KW
Kang
Sunghyouk
S
Park
D000085:Acetates / Q000378:metabolism; D000409:Alanine / Q000378:metabolism; D000970:Antineoplastic Agents / Q000494:pharmacology*; D056945:Hep G2 Cells; D022781:Hepatocytes / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism*; D006801:Humans; D049934:Isocoumarins / Q000494:pharmacology*; D007770:L-Lactate Dehydrogenase / Q000037:antagonists & inhibitors; D055442:Metabolome / Q000187:drug effects*; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D019289:Pyruvic Acid / Q000378:metabolism
D000085:Acetates; D000970:Antineoplastic Agents; D049934:Isocoumarins; C572336:galloflavin; D019289:Pyruvic Acid; D007770:L-Lactate Dehydrogenase; D000409:Alanine
NMR spectroscopy; cancer; galloflavin; metabolomics