Energy Management by Enhanced Glycolysis in G1-phase in Human Colon Cancer Cells In Vitro and In Vivo

Abstract

Activation of aerobic glycolysis in cancer cells is well known as the Warburg effect, although its relation to cellcycle progression remains unknown. In this study, human colon cancer cells were labeled with a cell-cycle phasedependent fluorescent marker Fucci to distinguish cells in G 1 -phase and those in S þ G 2 /M phases. Fucci-labeled cells served as splenic xenograft transplants in super-immunodeficient NOG mice and exhibited multiple metastases in the livers, frozen sections of which were analyzed by semiquantitative microscopic imaging mass spectrometry. Results showed that cells in G 1 -phase exhibited higher concentrations of ATP, NADH, and UDP-N-acetylglucosamine than those in S and G 2 -M phases, suggesting accelerated glycolysis in G 1 -phase cells in vivo. Quantitative determination of metabolites in cells synchronized in S, G 2 -M, and G 1 phases suggested that efflux of lactate was elevated significantly in G 1 -phase. By contrast, ATP production in G 2 -M was highly dependent on mitochondrial respiration, whereas cells in S-phase mostly exhibited an intermediary energy metabolism between G 1 and G 2 -M phases. Isogenic cells carrying a p53-null mutation appeared more active in glycolysis throughout the cell cycle than wild-type cells. Thus, as the cell cycle progressed from G 2 -M to G 1 phases, the dependency of energy production on glycolysis was increased while the mitochondrial energy production was reciprocally decreased.Implications: These results shed light on distinct features of the phase-specific phenotypes of metabolic systems in cancer cells. Mol Cancer Res; 11(9); 973-85. Ó2013 AACR.

Identification of N-acylhydrazone derivatives as novel lactate dehydrogenase A inhibitors

“…A recent investigation concerning the relationships between cell cycle progression and energy metabolism in cancer cells showed that the ATP requirement for G1 and S phases is largely met by accelerated glycolysis, while the energetic needs for G2/M phase are mainly derived from mitochondrial oxidative phosphorylation. On the basis of the published reports [37], our flow cytometry data, indicating a reduction of the cell population entering S phase and an increased fraction of G2/M cells, are compatible with effects exerted at the glycolytic level and can be a further evidence of the LDH-A inhibiting activity of compound 1. To assess whether the effect of compound 1 was not only limited to cell growth arrest, we measured by Western blot the expression of apoptosis markers (Fig.…”

Targeting Oxygen-Sensing Prolyl Hydroxylase for Metformin-Associated Lactic Acidosis Treatment

“…Photomicrographs were obtained using a BZ-9000 fluorescence microscope (Keyence, Osaka, Japan) at a magnification of ϫ20. Matrix-assisted laser desorption ionization-imaging mass spectrometry (MALDI-IMS) analysis of 2,8-dihydroxyadenine (m/z 168.05) in the kidneys of mice receiving a normal diet or an adenine-containing diet was performed in the positive ion mode using an atmospheric pressure MALDI-quadrupole ion trap time of flight mass spectrometer (MALDI-QIT-TOF-MS) (Shimadzu, Kyoto, Japan) as described previously (6,42,43).…”

Inhibition of the oxygen sensor PHD2 in the liver improves survival in lactic acidosis by activating the Cori cycle

“…Plasma samples were further purified by methanol-chloroform, as previously reported (31). 13 C-labeled glucose ( 13 C 1 -13 C 6 ), which is newly synthesized from administered 13 C 3 -lactate, was measured using the Nexera UHPLC (Ultra High Performance Liquid Chromatograph) system coupled with a LCMS-8030 triple quadrupole mass spectrometer (Shimadzu).…”

“…Extracted metabolites from the hepatocytes were analyzed using Agilent CE Capillary Electrophoresis System equipped with an Agilent 1100 series MSD mass spectrometer, as previously reported (31,32). Quantification of the metabolites was normalized with protein amount of the samples (μmol/g protein).…”