TWEAK诱导巨噬细胞源性外泌体miRNA抑制膀胱癌转移的研究  

TWEAK-stimulated macrophages inhibit metastasis of bladder cancer via exosomal microRNAs

作　　者：郑祖萍 夏先如[2] 周发友 唐晓磊 ZHENG Zu-ping;XIA Xian-ru;ZHOU Fa-you;TANG Xiao-lei(Department of Clinical Laboratory,Wudang Branch Hospital of Shiyan Taihe Hospital,Shiyan Hubei 442700;Department of Clinical Laboratory,Affiliated Taihe Hospital of Hubei University of Medicine,Shiyan Hubei 442000;Department of Urology Surgery,The Second Affiliated Hospital of Wannan Medical College,Wuhu Anhui 241000,China;Vascular Disease Research Center&Basic Medical Laboratory,The Second Affiliated Hospital of Wannan Medical College,Wuhu Anhui 241000,China)

机构地区：[1]湖北省十堰市太和医院武当山院区检验科,442700 [2]湖北省十堰市太和医院检验科,442000 [3]皖南医学院第二附属医院泌尿外科,安徽芜湖241000 [4]皖南医学院第二附属医院基础实验室&血管性疾病研究中心,安徽芜湖241000

出　　处：《蚌埠医学院学报》2022年第2期146-151,共6页Journal of Bengbu Medical College

基　　金：国家自然科学基金项目(81601806);安徽省教育厅高校科学研究重点项目(KJ2020A0614)。

摘　　要：目的:研究肿瘤坏死因子样弱凋亡诱导因子(tumor necrosis factor-like weak inducer of apoptosis,TWEAK)诱导的肿瘤相关巨噬细胞(TMs)源性的外泌体对膀胱癌细胞的转移影响。方法:使用TWEAK刺激THP-1诱导分化的TMs,检测其外泌体的特征。并使用miRNAs芯片检测TWEAK诱导的TMs中miRNAs的水平变化;随后通过激光共聚焦显微镜观察外泌体被膀胱癌ATCC 5637细胞系内化以及内化后ATCC 5637细胞中miRNAs的水平;同时检测外泌体处理对细胞的侵袭功能的影响以及活化的信号通路。结果:TWEAK不仅增加巨噬细胞及其分泌的外泌体中miRNA-17-3p的水平,而且还导致受体ATCC 5637细胞中miRNA-17-3p的水平升高,这最终降低了表皮生长因子受体(epidermal growth factor recepto,EGFR)/苏氨酸激酶(threonine kinase,AKT)/细胞外调节蛋白激酶(extracellular regulated protein kinase,ERK1)1/2途径的活性。在TMs中预先转染antagomiR-17-3p显著性降低了巨噬细胞、巨噬细胞分泌的外泌体和受体ATCC 5637细胞中miR-17-3p的水平,并伴随着ATCC 5637的转移增强,表明TMs源性的外泌体miR-17-3p参与了ATCC 5637细胞转移的调节,且通过抑制EGFR/AKT/ERK1/2途径得以实现。结论:TWEAK通过TMs将外泌体miRNAs转移到膀胱癌细胞中来抑制EGFR/AKT/ERK1/2途径,从而导致膀胱癌细胞的转移受到抑制。Objective:To explore the effect of exosomal-miRNAs from tumor necrosis factor-like weak inducer of apoptosis(TWEAK)-stimulated macrophages(TMs)on the metastasis of bladder cancer cells.Methods:The characteristics of exosomes derived from TWEAK-stimulated TMs,THP-1 induced differentiation were observed.Through the miRNAs microarray analysis,the expressions of miRNAs in exosomes derived from TMs treated with or without TWEAK were detected.The internalization of bladder cancer ATCC5637 cells and miRNAs after the internalization were detected by the laser scanning confocal microscope.The effect of exosome from TWEAK-stimulated TMs on metastasis and signal-pathway in ATCC5637 cells were detected.Results:TWEAK not only increased the levels of miR-17-3p in TMs and the secreted exosomes,but also resulted in an elevated levels of miR-17-3p in recipient ATCC 5637 cells,which eventually reduced the activity of the EGFR/AKT/ERK 1/2 pathway.Pre-transfection of antagomiR-17-3p in TMs substantially decreased the levels of miR-17-3p in macrophages,its secreted exosomes and the recipient ATCC 5637 cells with a concomitant enhancement of ATCC 5637metastasis,suggesting an involvement of exosomal miR-17-3p from TMs in modulating the metastasis of ATCC 5637cells.And miR-17-3p inhibited the metastasis of bladder cancer cells via the EGFR/AKT/ERK 1/2 pathway.Conclusions:TWEAK transfers exosomal miRNAs to bladder cancer cells via TMs to inhibit the EGFR/AKT/ERK 1/2 pathway,resulting in the inhibition of metastasis in bladder cancer cells.

关 键 词：膀胱肿瘤 肿瘤坏死因子样弱凋亡诱导因子 巨噬细胞 外泌体 微小RNA-17-3p 转移 

分 类 号：R737.14[医药卫生—肿瘤]