Objective To investigate the effects and mechanisms of PI3K/Akt on glycolysis,proliferation and migra-tion of human hepatocellular carcinoma HepG2 cells by regulating HKDC1.Methods Log-phase human hepatocellular carcinoma HepG2 cells cultured in vitro were divided into negative control group,LY294002 group and MK-2206 group,and the expression level of HKDC1 mRNA of cells in each group was detected by RT-qPCR,and the expression level of HKDC1 protein of cells in each group was analyzed by western blotting.The cells were divided into si-NC group(transfected with si-NC)and si-HKDC1 group(transfected with si-HKDC1),and the expression level of HKDC1 pro-tein in each group was analyzed by western blotting,the proliferation ability of cells in each group was detected by CCK-8 and EdU assays,the migration ability of cells in each group was detected by Wound healling and Transwell as-says,the extracellular acidification rate(ECAR)assay experiment to analyze the glycolysis and glycolytic capacity of cells in each group,and glucose and lactate content assay experiment to detect the intracellular glucose and lactate con-tent in each group.The cells were divided into control group,LY294002 group,overexpression of HKDC1+LY294002 group,MK-2206 group,and overexpression of HKDC1+MK-2206 group,and intracellular glucose and lactate contents were measured by glucose and lactate content assay.Results Compared with the control group,cells in the LY294002 group showed significantly lower HKDC1 mRNA expression(P＜0.001)and lower HKDC1 protein expression(P＜0.01);cells in the MK-2206 group showed lower HKDC1 mRNA expression(P＜0.01)and significantly lower HK-DC1 protein expression(P＜0.001).Compared with the si-NC group,cellular HKDC1 protein expression was reduced in the si-HKDC1 group(P＜0.001);compared with the si-NC group,cellular proliferation viability was reduced in the si-HKDC1 group(P＜0.05),and the number of EdU-positive cells was significantly reduced(P＜0.01);compared with the si-NC group,cellular wound healing rate was significantly reduced in the si-HKDC1 group(P＜0.001),and the number of migrating cells was significantly reduced(P＜0.001);cellular glycolysis and glycolytic capacity were signifi-cantly weakened in the si-HKDC1 group compared with the si-NC group(P＜0.01);and intracellular glucose and lactic acid content was significantly reduced in the si-HKDC1 group compared with the si-NC group(P＜0.05).The intracellular glucose and lactate contents were significantly higher in the overexpression HKDC1+LY294002 group compared with the LY294002 group(P＜0.01);and the intracellular glucose and lactate contents were significantly higher in the overexpression HKDC1+MK-2206 group compared with the MK-2206 group(P＜0.001,P＜0.01).Conclusion The PI3K/Akt signaling pathway promotes glycolysis,proliferation and migration of human hepatocellular carcinoma HepG2 cells via HKDC1.
关键词
磷脂酰肌醇-3-激酶/蛋白激酶B/含己糖激酶结构域的蛋白1/有氧糖酵解/增殖/迁移
Key words
phosphatidylinositol 3 kinase/protein kinase B/hexokinase domain containing protein 1/aerobic glycoly-sis/proliferation/migration
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