内源性β干扰素维持M1型巨噬细胞极化状态抑制肝癌细胞增殖和侵袭  

Endogenous IFN-β maintains M1 polarization status and inhibits proliferation and invasion of hepatocellular carcinoma cells

作　　者：谢昌利 郭变琴[1] 刘翠颖[1] 林艳[1] 吴碧涛 王秦[1] 李紫微[1] 涂植光[1] 

机构地区：[1]重庆医科大学检验医学院,临床检验诊断学教育部重点实验室,重庆400016 [2]重庆市璧山区人民医院检验科,重庆402760

出　　处：《细胞与分子免疫学杂志》2016年第7期865-869,875,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81172016)

摘　　要：目的研究内源性β干扰素(IFN-β)对M1型巨噬细胞极化状态及M1型巨噬细胞介导抗肝癌细胞增殖和侵袭的影响。方法体外建立单核系肿瘤细胞U937来源的M1巨噬细胞U937-M1模型,用实时定量PCR、ELISA及流式细胞术鉴定其表型。通过干扰IFN-β1基因或用抗体中和IFN-β,检测M1/M2巨噬细胞表型标志物表达的变化;用实时定量PCR和Western blot法检测干扰素调节因子1(IRF1)及IRF5的表达。收集不同处理组的U937-M1细胞培养上清为条件培养基(CM),分别培养Hep G2细胞及SMMC-7721细胞,用CCK-8法和TranswellTM侵袭实验分别检测CM对肝癌细胞增殖和侵袭的影响。结果与未激活型U937-M0细胞相比,U937-M1细胞白细胞介素12p35(IL-12p35)、IL-12p40、IL-12p70、IL-23p19、IL-6、肿瘤坏死因子α(TNF-α)和CD86表达显著增强;而两组均低表达CD206。阻断內源IFN-β作用后,与U937-M1细胞相比,上述M1型巨噬细胞相关表型标志物表达明显减弱;反之,M2型巨噬细胞表型标志物IL-10和CD206表达增强;且IRF1及IRF5表达均减弱;阻断IFN-β作用后,M1型巨噬细胞对肝癌细胞增殖及侵袭的作用从抑制转变为促进。结论阻断內源IFN-β后,M1型巨噬细胞极化状态以及U937-M1介导抗肝癌效应受到抑制;IFN-β可能参与调节IRF1和IRF5的表达,维持M1型巨噬细胞极化状态和功能。Objective To investigate the effect of endogenous interferon β （IFN-β） on the polarization of M1 macrophages as well as the proliferation and invasion activities of hepatocellular carcinoma cells （HCCs） mediated by M1 macrophages. Methods U937-M1 macrophages derived from human monocytic tumor cells U937 was established and the cell phenotypes were identified by real-time quantitative PCR, ELISA and flow cytometry. After IFN-β gene was knocked down with siRNA or IFN-β was neutralized with IFN-β monoantibody in U937-Ml macrophages, the change of MI/M2 phenotype was again analyzed by the above methods. The expressions of interferon regulatory factor 1 （IRF1） and IRF5 were detected by real-time quantitative PCR and Western blotting. The proliferation and invasion activities of HCCs, which were cultured with conditioned medium （CM） collected from different macrophage groups, were analyzed by CCK-8 assay and TranswellTMexperiments, respectively. Results U93？-M1 macrophages showed higher expressions of interleukin 12p35 （ IL-12p35 ）, interleukin 12p40 （ IL-12p40）, interleukin 12p70 （IL-12p70）, interleukin 23p19 （IL-23p19）, interleukin 6 （IL-6）, tumor necrosis factor α （TNF-α） and CD86 than U937-M0 did. But both U937-MO macrophages and U937-M! macrophages showed low expression of CD206. However, compared with the U937-M1 macrophages, the IFN-β-blocked U937-M1 macrophages presented decreased expressions of the above M1 macrophages-associated markers, but increased expressions of M2 macrophages-associated markers IL-10 and CD206, as well as lower expressions of IRF1 and IRFS. The inhibited proliferation/invasion activities of HCCs mediated by U937-M1 macrophages were reversed by IFN-β-blocked U937-M1 macrophages. Conclusion Blocking endogenous IFN-β could inhibit the U937-M1 polarization status and U937-Ml macrophages-mediated anti-tumor activity of HCCs. IFN-β might be involved in modulating the expressions of IRF1 and IRF5 as well as maintaining the M1 polarization sta

关 键 词：β干扰素(IFN-β) 巨噬细胞 极化 增殖 侵袭 干扰素调节因子(IRF) 

分 类 号：R392.12[医药卫生—免疫学] Q279[医药卫生—基础医学]