M2型巨噬细胞来源外泌体调控HGF/c-Met通路参与多发性骨髓瘤细胞转移的作用研究  

Effect of M2-type Macrophage-derived Exosomes on Multiple Myeloma Cell Metastasis by Regulation of HGF/c-Met Pathway

作　　者：木克热木·艾克帕尔 维尼拉·吐尔洪[1] 巴哈古丽·玉素甫 艾克拜尔·阿布都热衣木[1] Mukeremu Aikepaer;Vinila Tuerhong;Bahaguli Yusufu;Aikebaier Abudureyimu(Department of Hematology,Kashgar First People’s Hospital,Kashgar,Xinjiang Uygur Autonomous Region,844000,China)

机构地区：[1]喀什地区第一人民医院血液内科,新疆维吾尔自治区喀什市844000

出　　处：《医学分子生物学杂志》2024年第6期537-543,共7页Journal of Medical Molecular Biology

基　　金：新疆维吾尔自治区自然科学基金(No.2023D01C311)。

摘　　要：目的探究M2型巨噬细胞来源的外泌体对人多发性骨髓瘤细胞转移的影响及机制。方法在体外将THP-1细胞诱导分化成M0型和M2型巨噬细胞,实时荧光定量聚合酶反应(RT-qPCR)检测诱导后M2型巨噬细胞标志物血红蛋白清道夫受体(CD163)、白细胞介素-10(IL-10)、精氨酸酶-1(Arg-1)、转化生长因子-β1(TGF-β1)的表达水平,分离M2型巨噬细胞来源的外泌体;将RPMI-8226细胞分为对照组、M0-Exos组、M2-Exos组,Transwell小室法检测细胞迁移数目与侵袭数目,蛋白质印迹检测细胞中N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)、E-钙黏附蛋白(E-cadherin)及转录因子Snail的表达水平,并检测肝细胞生长因子(HGF)/c-Met相关蛋白的表达水平;接着将RPMI-8226细胞分为对照组、M2-Exos组、SU11274组、SU11274+M2-Exos组,Transwell小室法检测细胞迁移数目与侵袭数目,蛋白质印迹检测细胞中N-cadherin、Vimentin、E-cadherin及Snail表达水平。结果与M0型巨噬细胞比较,M2型巨噬细胞中CD163、IL-10、Arg-1、TGF-β1的mRNA表达水平均显著上调(P<0.05);从M2型巨噬细胞中分离的颗粒物中有CD9、CD63、TSG101、ALIX蛋白表达,由此判定该颗粒物为外泌体。与对照组比较,M2-Exos组细胞迁移与侵袭数目均显著增加(P<0.05),N-cadherin、Vimentin、Snail蛋白表达显著增加(P<0.05),E-cadherin蛋白表达显著减少(P<0.05),HGF蛋白表达及p-c-Met/c-Met比值也显著增加(P<0.05)。与M2-Exos组比较,SU11274+M2-Exos组细胞迁移与侵袭数目显著减少(P<0.05),N-cadherin、Vimentin、Snail蛋白表达显著减少(P<0.05),且E-cadherin蛋白表达显著增加(P<0.05)。结论M2型巨噬细胞来源的外泌体能够促进人多发性骨髓瘤细胞迁移、侵袭及EMT,加剧肿瘤细胞转移,该作用与调控HGF/c-Met通路有关。Objective To investigate the effect of M2-type macrophage-derived exosomes on human multiple myeloma cell metastasis and its mechanism.Methods THP-1 cells were induced to differentiate into M0 and M2 macrophages in vitro.Real-time fluorescent quantitative polymerase reaction(RT-qPCR)was used to detect the expression levels of hemoglobin scavenger receptor(CD163),interleukin-10(IL-10),arginase-1(Arg-1)and transforming growth factor-β1(TGF-β1)in induced M2 macrophages.Exosomes derived from M2 macrophages were isolated.RPMI-8226 cells were divided into 3 groups:control group,M0-Exos group and M2-Exos group.Then the cell migration and invasion were detected by Transwell.The expression levels of Ncadherin,Vimentin,E-cadherin and the transcription factor Snail,and hepatocyte growth factor(HGF)/c-Met related proteins were detected by Western blotting.Moreover,RPMI-8226 cells were then divided into 4 groups:control group,M2-Exos group,SU11274 group and SU11274+M2-Exos group,and cell migration and invasion were detected by Transwell,and the expression levels of N-cadherin,Vimentin,E-cadherin and Snail were detected by Western blotting.Results The mRNA expression levels of CD163,IL-10,Arg-1 and TGF-β1 in the M2 macrophages were significantly up-regulated when compared with those in the M0 macrophages(P<0.05).The particles isolated from the M2 macrophages showed protein expressions of CD9,CD63,TSG101 and ALIX,and were identified as exosomes.The numbers of cell migration and invasion in the M2-Exos group were significantly increased when compared with those in the control group(P<0.05),the protein expression levels of N-cadherin,Vimentin and Snail,and the HGF protein expression and p-c-Met/c-Met ratio were significantly increased(P<0.05),and the protein expression level of Ecadherin was significantly decreased(P<0.05).The numbers of cell migration and invasion in the SU11274+M2-Exos group were significantly decreased when compared with those in the M2-Exos group(P<0.05),the protein expression levels of N-cadherin,Vimentin

关 键 词：人多发性骨髓瘤 肿瘤相关巨噬细胞 外泌体 HGF/c-Met通路 

分 类 号：R733.3[医药卫生—肿瘤]