GPC3对肝癌细胞糖酵解的调控作用研究  

Effect of GPC3 on the Regulation of Glycolysis in Hepatocellular Carcinoma

作　　者：姚戈冰 张利旺 阴继凯 王成果 鲁建国 袁军 YAO Ge-bing;ZHANG Li-wang;YIN Ji-kai;WANG Cheng-guo;LU Jian-guo;YUAN Jun(Department of general surgery,Tangdu Hospital,The Air Force Military Medical university,Xi＇an,Shaanxi,710038,China;Department of medical administration,Tangdu Hospital,The Air Force Military Medical university,Xi＇an,Shaanxi,710038,China;Tangdu Hospital,The Air Force Military Medical university,Xi＇un,Shaanxi,710038,China)

机构地区：[1]空军军医大学唐都医院普外科,陕西西安710038 [2]空军军医大学唐都医院医教部,陕西西安710038 [3]空军军医大学唐都医院,陕西西安710038

出　　处：《现代生物医学进展》2018年第10期1860-1864,共5页Progress in Modern Biomedicine

基　　金：陕西省社会发展科技攻关项目(2011k13-01-01;2012k13-02-13)

摘　　要：目的:探讨GPC3(glypican 3)在肝癌细胞糖酵解中的调控作用。方法:采用si RNA(small interfering RNA)干扰肝癌细胞中GPC3的表达后,采用q PCR(quantitative PCR)与Western blot实验检测肿瘤糖酵解关键调控分子Glut1(glucose transporter-1)、HK2(hexokinase 2)与LDH-A(Lactate Dehydrogenase A)的表达,通过检测培养液中葡萄糖的减少量分析GPC3对细胞葡萄糖摄取情况,通过检测培养液中乳酸含量与PH值分析GPC3对细胞乳酸产生的影响,通过检测细胞的氧耗速率,分析GPC3对线粒体氧化磷酸化功能的影响。结果:干扰肝癌细胞中GPC3的表达可抑制糖酵解关键调控分子Glut1、HK2与LDH-A表达,降低肝癌细胞葡萄糖摄取速率和细胞氧耗速率,且细胞培养液PH升高,乳酸产生减少。结论:肝癌细胞中GPC3高表达通过上调糖酵解关键调控分子Glut1、HK2与LDH-A表达而促进肝癌细胞糖酵解效应,同时抑制线粒体氧化磷酸化活性。这些结果进一步提示糖代谢重编程可能是GPC3促进肝癌增殖与转移的重要机制。Objective： To investigate the role of GPC3（glypican 3） in the regulation of glycolysis in HCC cells.Methods： The expression of Glut1（glucose transporter-1）,HK2（hexokinase 2） and LDH-A（Lactate Dehydrogenase A） were detected by q PCR and Western blot analysis after the GPC3 was knocked-down by si RNA in HCC cells.Glucose uptake was analyzed by detecting the reduction of glucose in the culture medium of HCC cells.Lactate production was analyzed by detecting the concentration of Lactate and p H value in the culture medium of HCC cells.Oxidative phosphorylation was analyzed by measuring the rate of oxygen consumption in HCC cells.Results： After GPC3 was knocked-down by si RNA in HCC cells,the expression of Glut1,HK2 and LDH-A,and glucose uptake were significantly decreased,while p H value of the culture medium and the rate of oxygen consumption was significantly increased.Conclusion： High expression of GPC3 can promote the glycolytic effect of HCC cells by upregulating the expression of Glut1,HK2 and LDH-A,which are the key regulators of glycolysis,and inhibit the oxidative phosphorylation activity of HCC cells.These results suggest that glucose metabolism reprogramming is crucial for GPC3 to promote the proliferation and metastasis of hepatocellular carcinoma.

关 键 词：GPC3 肝癌 糖酵解 氧化磷酸化 

分 类 号：R-33[医药卫生] R735.7