探讨髓系白血病细胞株糖酵解表型特征  

Characterization of Glycolytic Phenotype of Myeloid Leukemic Cell Lines

作　　者：陈莎娜[1] 鲜敬荣[1] 王娟[1] 贺金刚[1] 全静[1] 张帅帅[1] 张伶[1] 

机构地区：[1]重庆医科大学检验医学院,临床检验诊断学教育部重点实验室,重庆市重点实验室,重庆400016

出　　处：《中国细胞生物学学报》2014年第1期39-47,共9页Chinese Journal of Cell Biology

基　　金：重庆市科委自然科学基金计划(批准号:cstc2013jcyjA10035)资助的课题~~

摘　　要：探讨髓系白血病细胞株的糖酵解表型特征及其潜在的调控机制。葡萄糖试剂盒和乳酸试剂盒分别检测5株白血病细胞培养上清液中的葡萄糖消耗(G)和乳酸生成含量(L),计算L/G比值来评估糖酵解水平;定量PCR检测糖酵解相关基因GLUT、MCT1 mRNA表达;CCK8法检测细胞体外增殖能力;Western blot检测AKT蛋白磷酸化水平。结果显示,KG1a和K562细胞体外培养24 h后的L/G比值分别为1.78和1.71,接近糖酵解表型时L/G为2的比值,同时这两株细胞高表达糖酵解相关基因GLUT1和MCT1 mRNA。低糖(0.5 mmol/L)、中糖(5 mmol/L)、高糖(10 mmol/L)处理KG1a和K562细胞40 h后,两株细胞的增殖能力、葡萄糖消耗和乳酸生成随葡萄糖浓度增加而增强,高糖组增加更为显著(P<0.05)。相反,若糖酵解抑制剂2-DG(0,5,10 mmol/L)处理白血病细胞40 h后,两株细胞的增殖能力及糖酵解代谢水平随2-DG浓度增加而降低,高浓度2-DG组(10 mmol/L)降低更为显著(P<0.05)。此外,AKT抑制剂低浓度(5μmol/L)短时间(12 h)处理后能抑制白血病细胞AKT蛋白磷酸化水平,同时降低细胞的葡萄糖消耗和乳酸生成(P<0.05)。该研究提示髓系白血病细胞具有高糖酵解表型,AKT可能参与调控白血病的糖代谢过程,这有助于阐明白血病的能量代谢特征以及为白血病的靶向抗代谢治疗奠定基础。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 capa- bility 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 KGla 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 KGla and K562 cells. The proliferation capacity, glucose consumption and lactate production in KGIa 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 glyco- lytic 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 le

关 键 词：髓系白血病 糖代谢 糖酵解 AKT 

分 类 号：R733.7[医药卫生—肿瘤]