胃癌细胞外泌体经miRNA触发肝Kupffer细胞M2型极化以促进肝转移前生态位的形成  

Exosomes derived from gastric cancer cells trigger M2 polarization of hepatic Kupffer cells via miRNA to promote the formation of pre-metastatic hepatic niche

作　　者：张轩 刘炜 付海啸 李腾腾 刘浩 符炜 王凯 Zhang Xuan;Liu Wei;Fu Haixiao;Li Tengteng;Liu Hao;Fu Wei;Wang Kai(Department of Gastrointestinal Surgery,the Affiliated Hospital of Xuzhou Medical University,Xuzhou 221002,China)

机构地区：[1]徐州医科大学附属医院胃肠外科,徐州221002

出　　处：《中华微生物学和免疫学杂志》2024年第9期762-770,共9页Chinese Journal of Microbiology and Immunology

基　　金：徐州市卫健委青年课题项目(XWKYHT20230057)。

摘　　要：目的探讨胃癌细胞来源的外泌体与肝Kupffer细胞间的相互作用对胃癌肝转移的影响,并揭示其潜在的作用机制。方法通过裸鼠模型、病毒转染和流式分选技术,从胃癌细胞株(MKN45)中构建具有高度肝转移潜能的细胞株(MKN45-HL)。超速离心法收集外泌体,并通过纳米流式仪、透射电镜以及Western blot等进行表征和鉴定。构建胃癌细胞外泌体训导的裸鼠肝转移模型,并经活体成像技术监测肝转移灶的发展。通过细胞培养、qRT-PCR和免疫荧光染色等分析胃癌细胞外泌体对巨噬细胞极化的作用。利用外泌体miRNA的组学分析和qRT-PCR筛选验证胃癌外泌体促巨噬细胞M2极化的miRNA分子靶点。结果胃癌来源的外泌体主要集中在肝脏,大多数被肝内巨噬细胞所摄取,且在体外培养和裸鼠体内都能促进巨噬细胞向M2型极化。MKN45与MKN45-HL外泌体训导的裸鼠经脾脏注射小鼠前胃癌细胞后均出现明显的肝转移灶,且MKN45-HL外泌体表现出更强的促肝脏巨噬细胞M2极化和促胃癌细胞肝转移的能力。两者的miRNA组学分析显示了很多差异性表达的miRNAs。高肝转移潜力的胃癌细胞系和肝转移患者血清外泌体中miR-519a-3p显著升高,可经外泌体被巨噬细胞内化,对胃癌肝转移有预测价值,并与肝转移预后密切相关。结论胃癌细胞来源的外泌体经miR-519a-3p触发肝Kupffer细胞的M2样极化,推进胃癌的肝转移进程,可能是塑造胃癌肝转移前生态位微环境的重要分子。本研究为揭示胃癌肝转移机制提供了新视角,并为未来的治疗策略提供了潜在靶点。ObjectiveTo investigate the influence of the interaction between gastric cancer(GC)cell-derived exosomes and hepatic Kupffer cells on GC with liver metastasis and analyze the potential mechanism.MethodsCells with high hepatic metastatic potential(MKN 45-HL)were constructed from a parental GC cell line(MKN 45)using a nude mouse model and methods of viral transfection and flow sorting.Exosomes were collected using ultra-centrifugation and characterized by electron microscopy,nanoparticle tracking system and Western blot.A nude mouse model of liver metastasis induced by GC cell-derived exosomes was constructed,and the development of liver metastases was monitored by live imaging.The regulatory effects of GC cell-derived exosomes on macrophage polarization were assessed by cell culture,qRT-PCR,and immunofluorescence staining.Using the omics analysis of exosomal miRNA and qRT-PCR,the molecular targets by which exosomes specifically promoting macrophage M2 polarization were screened and validated.ResultsGC cell-derived exosomes were mainly concentrated in the liver,most of which were ingested by intrahepatic macrophages,and could promote macrophages to M2 polarization in both in vitro culture and nude mice.Both groups of mice trained with MKN 45 and MKN 45-HL exosomes showed obvious liver metastases after mouse forestomach carcinoma(MFC)cells injection through the spleen,and MKN 45-HL exosomes showed a much stronger ability to promote hepatic macrophage M2 polarization and liver metastasis of MFC cells.Moreover,the miRNA omics analysis revealed a lot of differentially expressed miRNAs between MKN 45-derived and MKN 45-HL-derived exosomes.The expression of miR-519a-3p increased significantly in the exosomes derived from MKN 45-HL cell line and the clinical serum of GC patients with liver metastasis.It was found that miR-519a-3p could be internalized by macrophages through exosomes delivery.Furthermore,the miR-519a-3p in exosomes from patient′s serum had a predictive value for GC with liver metastasis and was closely a

关 键 词：胃癌 外泌体 KUPFFER细胞 M2样极化 肝转移前生态位 

分 类 号：R735.2[医药卫生—肿瘤]