MAFB对肿瘤相关巨噬细胞极化及功能的影响  

Impact of MAFB on polarization and function of tumor associated macrophages

作　　者：陈笑天 陈翀[1] 罗云萍[1] CHEN Xiaotian;CHEN Chong;LUO Yunping(Department of Immunology,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免疫学系,北京100005

出　　处：《基础医学与临床》2024年第7期965-973,共9页Basic and Clinical Medicine

基　　金：国家自然科学基金(82273220,82373100);常州西太湖细胞前沿技术发展基金会重点培育项目(2022-P009)。

摘　　要：目的探究MAFB转录因子对实体肿瘤中巨噬细胞极化表型和功能的影响。方法通过Kaplan-Meier数据库的RNA-seq数据分析MAFB与乳腺癌和肺腺癌患者生存期的相关性;流式细胞测量术检测原代人单核细胞中MAFB的表达;RT-qPCR和Western blot检测人单核细胞白血病细胞系(THP-1)中敲低MAFB的表达水平;利用巨噬细胞体外极化、细胞共培养、RT-qPCR和流式细胞测量术检测巨噬细胞的极化表型;吞噬实验测定巨噬细胞的吞噬能力;体外杀伤实验测定巨噬细胞的肿瘤杀伤能力。结果与MAFB低表达组相比,MAFB高表达乳腺癌或肺腺癌患者生存期降低(P<0.05);蛋白质互作网络分析显示MAFB蛋白与巨噬细胞极化蛋白有相关性;与其他类型的组织细胞相比,人单核-巨噬细胞特异性高表达MAFB基因;佛波酯(PMA)诱导THP-1细胞分化后MAFB表达显著上调(P<0.05);在THP-1中敲低MAFB可以抑制M2相关基因表达(P<0.05);与肿瘤细胞共培养后敲低MAFB的巨噬细胞抑制其极化基因表达(P<0.05);敲低MAFB不影响巨噬细胞对肿瘤细胞的吞噬能力(P>0.05);敲低MAFB增强巨噬细胞体外肿瘤杀伤能力(P<0.05)。结论本研究初步证实在实体肿瘤中MAFB转录因子具有调控巨噬细胞的M2极化作用和抗肿瘤功能。Objective To investigate the impact of the MAFB transcription factor on macrophage polarization phe⁃notype and function.Methods Analysis of correlation of MAFB expression with survival period of patients of breast cancer and lung adenocarcinoma was performed by RNA⁃seq data from the Kaplan⁃Meier database.Flow cytometry was applied to detect MAFB expression in primary human monocytes.Expression of MAFB mRNA or protein of hu⁃man acute monocytic leukaemia cell line(THP⁃1)was detected by RT⁃qPCR or Western blot.Detection of macro⁃phages phenotypes was performed by macrophages polarization in vitro,co⁃culture,RT⁃qPCR and flow cytometry.Phagocytosis assay was used to assess changes in phagocytic ability of macrophages.In vitro killing assay to evaluate changes in tumor⁃killing ability of macrophages.Results Compared to the MAFB low⁃expression group,patients with high MAFB expression in breast cancer and lung adenocarcinoma exhibited a decreased survival period(P<0.05).Protein interaction networks showed a correlation between MAFB protein and macrophage polarization proteins.Compared to other cell type,MAFB was specifically highly expressed in human monocyte⁃macrophage cells.Phorbol myristate ester(PMA)induced an increased MAFB expression in THP⁃1 cells(P<0.05).Knockdown of MAFB inhibited the expression of M2⁃related genes(P<0.05).Co⁃culture with tumor cells and knockdown of MAFB suppressed expression of polarization gene(P<0.05).Knockdown of MAFB did not affect the phagocytic abilityP>0.05)but enhanced tumor⁃killing ability of macrophages(P<0.05).Conclusions This investigation proves potential function of MAFB transcription factor in regulation of macrophage M2 polarization and anti⁃tumor effect in solid tumors.

关 键 词：MAFB 巨噬细胞 M1/M2极化 吞噬作用 肿瘤杀伤效应 

分 类 号：R392.12[医药卫生—免疫学]