TIPIN分子通过调控糖代谢促进肝癌细胞生长  

TIPIN promotes hepatocellular carcinoma cell growth by regulating glucose metabolism

作　　者：卫宣均 刘文翰 李莉[3] 关琳[4] WEI Xuanjun;LIU Wenhan;LI Li;GUAN Lin(Second Clinical College of China Medical University,Shenyang 110004;Department of Clinical Laboratory,Shengjing Hospital Affiliated to China Medical University,Shenyang 110004;Department of Scientific Research and Discipline,First Affiliated Hospital of China Medical University,Shenyang 110002;Department of Gastroenterology,First Affiliated Hospital of China Medical University,Shenyang 110002,China)

机构地区：[1]中国医科大学第二临床学院,沈阳110004 [2]中国医科大学附属盛京医院检验科,沈阳110004 [3]中国医科大学附属第一医院科研与学科部,沈阳110002 [4]中国医科大学附属第一医院消化内科,沈阳110002

出　　处：《临床与病理杂志》2023年第11期1928-1936,共9页Journal of Clinical and Pathological Research

摘　　要：目的:永恒昼夜节律相互作用蛋白(timeless circadian clock interacting protein,TIPIN)在肝癌中高表达且与肝癌的恶性进展有关,但TIPIN在肝癌中的生物学作用尚不清楚。本研究探讨TIPIN对肝癌细胞糖代谢的影响及其分子机制。方法:在基因表达谱数据交互分析(the Gene Expression Profiling Interactive Analysis,GEPIA)数据库中分析TIPIN在肝癌组织中的表达及其与患者预后的关系。实时定量PCR和蛋白质印迹实验检测肝癌细胞和正常肝细胞中TIPIN的表达,在高表达TIPIN的MHCC97H和低表达TIPIN的MHCC97L中,分别敲减TIPIN和过表达TIPIN,检测细胞葡萄糖摄取、乳酸生成、pH、细胞生长、氧耗量和糖代谢关键调控分子的表达。结果:TIPIN在肝癌组织中的表达高于正常肝组织(P<0.05),且高表达TIPIN的患者总生存期及无病生存期较低表达TIPIN的患者短(均P<0.05)。与正常肝细胞HL7702相比,肝癌细胞MHCC97L、HepG2、HuH7、MHCC97H中TIPIN的表达水平均明显升高(均P<0.01)。TIPIN敲减后细胞葡萄糖摄取和乳酸生成水平降低,而细胞耗氧量和培养基p H增加(均P<0.05);TIPIN过表达后细胞葡萄糖摄取和乳酸生成水平升高,而细胞耗氧量和培养基pH降低(均P<0.05)。TIPIN敲减后p53表达上调(P<0.01),过表达TIPIN后p53表达减少(P<0.01),而缺氧诱导因子1α(hypoxia inducible factor-1α,HIF-1α)和髓细胞增生原癌基因(myelocytomatosis oncogene,c-MYC)均未见改变。在MHCC97H细胞中干扰p53后可以恢复只敲减TIPIN造成的生长抑制(P<0.01),且糖代谢表型与对照组比较差异无统计学意义(P>0.05);在MHCC97L细胞中过表达TIPIN后细胞生长能力增强(P<0.01),但过表达TIPIN和p53会抑制细胞生长(P<0.01),且糖代谢表型与对照组比较差异无统计学意义(P>0.05);结论:TIPIN通过抑制p53的表达来增强细胞糖酵解和抑制氧化磷酸化,进而促进肝癌细胞生长。Objective:The timeless circadian clock interacting protein(TIPIN)is highly expressed in hepatocellular carcinoma and is associated with the malignant progression of hepatocellular carcinoma.However,the biological role of TIPIN in hepatocellular carcinoma remains unclear.This study aims to investigate the impact of TIPIN on glucose metabolism in hepatocellular carcinoma cells and explore the molecular mechanisms involved.Methods:The expression of TIPIN in hepatocellular carcinoma tissues and its relationship with patient prognosis were analyzed using the Gene Expression Profiling Interactive Analysis(GEPIA)database.The expression of TIPIN in hepatocellular carcinoma cells and normal liver cells was detected by Western blotting and real-time RT-PCR.TIPIN was knocked down in MHCC97H cells with high TIPIN expression and overexpressed in MHCC97L cells with low TIPIN expression.Glucose uptake,lactate production,pH,cell growth,oxygen consumption,and the expression of key molecules in glucose metabolism were measured.Results:TIPIN expression was higher in hepatocellular carcinoma tissues than in normal liver tissues(P<0.05),and patients with high TIPIN expression had shorter overall survival and disease-free survival compared to those with low TIPIN expression(P<0.05).Compared to the normal liver cell line HL7702,the expression of TIPIN was significantly increased in the liver cancer cell lines MHCC97L,HepG2,HuH7,and MHCC97H(all P<0.01).After TIPIN knockdown,glucose uptake and lactate production decreased,while oxygen consumption and pH increased(all P<0.05).Conversely,TIPIN overexpression led to increased glucose uptake and lactate production,along with decreased oxygen consumption and pH(P<0.05).TIPIN knockdown increased p53 expression(P<0.01),while TIPIN overexpression decreased p53 expression(P<0.01);however,hypoxiainducible factor-1α(HIF-1α)and myelocytomatosis oncogene(c-MYC)expression did not change.In MHCC97H cells,interference with p53 expression restored the growth inhibition caused by TIPIN knockdown(P<0.01)

关 键 词：永恒昼夜节律相互作用蛋白 肝癌 糖代谢 生长 

分 类 号：R735.7[医药卫生—肿瘤]