Objective To explore the role of cytidine/uridine monophosphate kinase 2( CMPK2) in the immune-mediated antitumor effect of IFNα in 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 overexpress-ing 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 overexpress-ing Huh7 cells. Results The transcription and protein level of CMPK2 were significantly enhanced after the treat-ment of IFNα for 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 IFNα increases the expression of CMPK2 in Huh7 cells to activate the expression of inflammatory cytokines in macrophages.%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-qPCR 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CellTiter-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-qPCR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.
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