IFNα上调人肝癌细胞系胞苷单磷酸激酶2表达激活小鼠巨噬细胞系  

IFNα induces cytidine/uridine monophosphate kinase 2 expression in hepatocarcinoma cell line to activate macrophages of mice

作　　者：成丽琴 张祝琴[1] 陈厚早[1] CHENG Li-qin;ZHANG Zhu-qin;CHEN Hou-zao(State Key Laboratory of Medical Molecular Biology,Institute of Basic Medical Sciences CAMS, School of Basic Medicine PUMC,Beijing 100005,China)

机构地区：[1]中国医学科学院基础医学研究所北京协和医学院基础学院医学分子生物学国家重点实验室,北京100005

出　　处：《基础医学与临床》2018年第7期944-949,共6页Basic and Clinical Medicine

基　　金：国家重点研发计划(2016YFC0903900);北京协和医学院协和青年科研基金(3332016047);中国医学科学院医学与健康科技创新工程(2016-I2M-1-011;2017-I2M-1-008)

摘　　要：目的探讨CMPK2在IFNα治疗肝癌中的抗肿瘤免疫反应作用。方法用RT-q PCR和Western blot检测IFNα处理后胞苷单磷酸激酶2(CMPK2)在肝癌细胞系Huh7中的表达。通过构建的稳定表达CMPK2的慢病毒感染Huh7细胞,用Cell Titer-Glo ATP荧光检测过表达CMPK2的Huh7细胞及其上清中ATP的水平。用RT-q PCR检测过表达CMPK2的Huh7细胞上清对巨噬细胞中炎性因子的表达的影响。结果 IFNα处理6 h后,Huh7细胞中CMPK2的转录水平和蛋白水平显著上调(P<0.01);CMPK2显著上调Huh7细胞及其上清中ATP的水平(P<0.01);用稳定过表达CMPK2的Huh7细胞上清处理巨噬细胞6 h后,巨噬细胞中IL1β、IL6和CCL5的转录水平明显高于对照组(P<0.01)。结论 IFNα上调Huh7细胞中CMPK2水平,激活巨噬细胞中炎性因子的表达。Objective To explore the role of cytidine/uridine monophosphate kinase 2(CMPK2)in the immune-mediated antitumor effect of IFNαin hepatocellular carcinoma.Methods RT-qPCR and Western blot were used to analyze the expression of CMPK2 in Huh7 after the treatment of IFNα.The CMPK2 overexpressing Huh7 cells were generated by stably infecting with lentivirus.The ATP level in the cells and the supernatant of CMPK2 overexpressing Huh7 cells were measured by CellTiter-Glo ATP fluorescence assay.RT-qPCR was applied to test the expression of inflammatory cytokines in macrophages under the treatment of the supernatant of CMPK2 overexpressing Huh7 cells.Results The transcription and protein level of CMPK2 were significantly enhanced after the treatment of IFNαfor 6 hours(P<0.01).CMPK2 increased the ATP level in the cells and supernatant of Huh7 cells(P<0.01).The supernatant of CMPK2 overexpressing Huh7 cells activated the expression of IL1β,IL6 and CCL5 in macrophages(P<0.01).Conclusions IFNαincreases the expression of CMPK2 in Huh7 cells to activate the expression of inflammatory cytokines in macrophages.

关 键 词：肝癌细胞 胞苷单磷酸激酶2 干扰素Α 

分 类 号：R735.7[医药卫生—肿瘤]