Characterization of Glycolytic Phenotype of Myeloid Leukemic Cell Lines

Abstract: To explore the glycolytic phenotype of myeloid leukemic cell lines and its potential mechanisms，the glucose consumption (G) and lactate production (L) in five leukemic cell lines were detected by glucose assay kit and lactate measurement kit， respectively. The glycolytic level was evaluated by the ratio of L/G; The glycolytic metabolism related genes (GLUT1， MCT1) were determined by quantitative Real-time PCR; Cell proliferation capability was analyzed by CCK-8 (cell counting kit-8) assay. Western blot was used to detect phosphorylation level of AKT protein. The results showed that the ratio of L/G was 1.78 for KG1a and 1.71 for K562， which was closer to 2 (if the glucose was metabolized only through glycolysis， L/G ratio should be 2) comparing to the other three leukemic cell lines. Additionally， the expressions of both GLUT1 and MCT1 mRNA were much higher in KG1a and K562 cells. The proliferation capacity， glucose consumption and lactate production in KG1a and K562 cells had enhanced after treated by different concentration of glucose (0.5， 5， 10 mmo/L) for 40 h， significantly higher in the high glu cose group (10 mmo/L) (P<0.05). However， the cell proliferation and the levels of glycolytic metabolism were notably decreased in a dose-dependent manner after treatment with 2-DG (0， 5， 10 mmol/L) for 40 h， significantly lower in high 2-DG group (10 mmo/L) (P<0.05). The low concentration of AKT IV (5 μmol/L) and short treatment time (12 h) was use to observe the metabolic change without the reduction of cell number. The results showed that AKT IV significantly reduced glucose consumption and lactate production， and inhibited the phosphorylation level of AKT in leukemic cells (P<0.05). These results demonstrate that myeloid leukemia cells exhibit the highly glycolytic phenotype， and AKT may involve in the regulation of glycolysis， which will contribute to clarify the leukemic cells’ characteristics of energy metabolism and facilitate the future leukemia therapy targeting metabolic pathways.

CSTR: 32200.14.cjcb.2014.01.0007