IRF1对M1巨噬细胞极化及其抗肿瘤效应的影响  

Effects of IRF1 on polarization and antitumor function of M1 microphage

作　　者：谢昌利 刘翠颖[1] 林艳[1] 吴碧涛 王秦[1] 李紫微[1] 涂植光[1] XIE Chang-li LIU Cui-ying LIN Yan WU Bi-tao WANG Qin LI Zi-wei TU Zhi-guang(Key Laboratory of Laboratory Medical Diagnostics of Education Ministry, College of Laboratory Medicine, Chongqing Medical University Chongqing 400016, China)

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试验室,重庆400016

出　　处：《基础医学与临床》2017年第2期189-196,共8页Basic and Clinical Medicine

基　　金：国家自然科学基金(81172016)

摘　　要：目的研究IRF1对M1巨噬细胞极化及M1介导的抗肝癌细胞增殖和凋亡的影响。方法构建单核细胞U937来源M1巨噬细胞模型(U937-M1),将细胞分为4组:用PMA诱导的未活化巨噬细胞组(M0),用PMA,IFN-γ和LPS处理的M1型巨噬细胞组(M1),用siRNA干扰IRF1的M1型巨噬细胞组(si IRF1)以及阴性干扰的M1型巨噬细胞组(si C)。用流式细胞术检测M1/M2特异表面标志物CD86/CD206的表达;q PCR检测M1/M2相关基因(IL-12p35,IL-12p40,IL-23p19,IL-6,TNF-α/IL-10)及IFNB1的表达;ELISA检测IL-12p70,IL-10及IFN-β的表达;Western blot检测IRF1及IRF5的表达;CCK8和流式细胞术分别检测Hep G2及SMMC-7721增殖和凋亡。结果与U937-M1组相比,干扰IRF的M1组CD86表达降低,但CD206升高(P<0.05);mRNA水平上,IL-12p35,IL-12p40,IL-23p19,IL-6,TNF-α以及IFNB1表达降低,但IL-10表达增高(P<0.01);蛋白水平上,IL-12p70,IFN-β及IRF5表达降低,但IL-10表达增高(P<0.05)。IRF1干扰后,M1巨噬细胞促进肝癌细胞增殖、抑制其凋亡(P<0.05)。结论干扰IRF1后,M1巨噬细胞极化状态受损,甚至部分向M2型转变;其抗肿瘤效应转变为促肿瘤效应;且IRF1可能参与调节IFN-β与IRF5的表达。Objective To study if IRF1 could regulate the polarization by IRF1 and M1 status and affect M1 mediated antitumor function. Methods U937 derived M1 macrophage（ U937-M1） model was established. The cells were devided into 4 groups： the PMA pretreated unpolarized macrophage（ M0）,the PMA,IFN-γ and LPS induced M1 macrophage（ M1）,the siRNA of IRF1 knocked down M1 macrophage（ si IRF1） and the negative control siRNA treated M1 macrophage（ si C）. Furthermore,the expression of CD86 and CD206 was detected by flow cytometry,the M1 / M2 associated markers（ IL-12p35,IL-12p40,IL-23p19,IL-6,TNF-α / IL-10） and IFNB1 were analyzed by q PCR,the expression of IL-12p70 and IL-10 was examined by ELISA,the expression of IRF1 and IRF5 was detected by Western blot,the proliferration and apoptosis of HCC were analyzed by CCK8 and flow cytometry,respectively. Results Compared with the U937-M1,the IRF1 knocked down group showed impaired CD86 expression,but enhanced CD206 expreesion（ P〈0. 05）; the expression of M1 related cytokines including IL-12p35,IL-12p40,IL-23p19,IL-6,TNF-α and IFNB1 was decreased,but M2 related cytokine IL-10 level was increased（ P〈0. 01）; the expression of IFN-β,IL-12p70 and IRF5 was impaired,but IL-10 was enhanced（ P〈0. 05）. In IRF1 knocked down U937-M1,the CCK8 analysis indicated that the M1 mediated anti-proliferation effects on hepatomacarcinoma cell were turned to pro-proliferation（ P〈0. 05）; the flow cytometry showed that the M1 mediated pro-apoptosis effects were reversed to anti-apoptosis（ P〈0. 01）. Interestingly,IRF5 and IFN-β were decreased at both mRNA and protein levels in IRF1 knocked down U937-M1 compared with the U937-M1（ P〈0. 01）. Conclusions IRF1 may partly modulate IRF5 and IFN-β,and further regulate M1 polarization and its antitumor effects.

关 键 词：IRF1 M1极化 抗肿瘤 IRF5 

分 类 号：R73-3[医药卫生—肿瘤]