Zhang Shaofeng, Li Danqing, Wang Haijun, Liu Bo, Du Fan, Wang Qing
Department of Thoracic surgery, Xingtai People's Hospital, Xingtai, China.
Department of Radiotherapy, Xingtai People's Hospital, Xingtai, China.
Cell Biochem Biophys. 2025 Mar;83(1):633-646. doi: 10.1007/s12013-024-01496-2. Epub 2024 Sep 5.
Cancer-associated fibroblasts (CAFs) represent one of the major components of the tumor stroma, which might create an immunosuppressive tumor microenvironment by inducing and functionally polarizing protumoral macrophages. Previous studies indicated that exosomes derived from CAFs might transmit regulating signals and boost esophageal squamous cell carcinoma (ESCC) development. This study is designed to explore the role and mechanism of CAFs-derived exosomal microRNA-889-3p (miR-889-3p) in ESCC progression. Macrophage polarization was detected using flow cytometry. miR-889-3p, Tumor necrosis factor alpha (TNF-α), and inducible nitric oxide synthase (iNOS) levels were detected by real-time quantitative polymerase chain reaction (RT-qPCR). Cell proliferation, cycle progression, migration, and invasion were assessed using Cell Counting Kit-8 (CCK-8), 5-ethynyl-2'-deoxyuridine (EdU), scratch assay, and Transwell assays. α-SMA, FAP, CD63, CD81, and signal transducer and activator of transcription 1 (STAT1) protein levels were detected using western blot. Exosomes were characterized using an electron microscope and nanoparticle tracking analysis (NTA). Binding between miR-889-3p and STAT1 was predicted by Starbase, and verified by a dual-luciferase reporter and RNA pull-down. The effect of CAFs-derived exosomal miR-889-3p on ESCC tumor growth in vivo was detected using mice xenograft assay. miR-889-3p level was decreased in LPS-induced M0 macrophages. CAF-derived exosomal miR-889-3p knockdown suppressed ESCC proliferation, migration, and invasion. CAFs might transfer miR-889-3p to M0 macrophages via exosomes. STAT1 was a target of miR-889-3p. Besides, in vivo studies confirmed that CAFs-derived exosomal miR-889-3p can accelerate ESCC tumor growth by regulating STAT1. CAFs-derived exosomal miR-889-3p facilitates esophageal squamous cell carcinoma cell proliferation, migration, and invasion by inhibiting M1 macrophage polarization through down-regulation of STAT1, providing a promising therapeutic target for ESCC.
癌症相关成纤维细胞（CAFs）是肿瘤基质的主要成分之一，它可能通过诱导肿瘤相关巨噬细胞并使其功能极化来创建一个免疫抑制性肿瘤微环境。先前的研究表明，源自CAFs的外泌体可能传递调节信号并促进食管鳞状细胞癌（ESCC）的发展。本研究旨在探讨源自CAFs的外泌体微小RNA-889-3p（miR-889-3p）在ESCC进展中的作用及机制。使用流式细胞术检测巨噬细胞极化。通过实时定量聚合酶链反应（RT-qPCR）检测miR-889-3p、肿瘤坏死因子α（TNF-α）和诱导型一氧化氮合酶（iNOS）的水平。使用细胞计数试剂盒-8（CCK-8）、5-乙炔基-2'-脱氧尿苷（EdU）、划痕试验和Transwell试验评估细胞增殖、细胞周期进程、迁移和侵袭。使用蛋白质印迹法检测α-平滑肌肌动蛋白（α-SMA）、成纤维细胞活化蛋白（FAP）、CD63、CD81和信号转导子和转录激活子1（STAT1）的蛋白水平。使用电子显微镜和纳米颗粒跟踪分析（NTA）对外泌体进行表征。通过Starbase预测miR-889-3p与STAT1之间的结合，并通过双荧光素酶报告基因和RNA下拉实验进行验证。使用小鼠异种移植试验检测源自CAFs的外泌体miR-889-3p对ESCC肿瘤在体内生长的影响。在脂多糖诱导的M0巨噬细胞中，miR-889-3p水平降低。源自CAF的外泌体miR-889-3p敲低可抑制ESCC的增殖、迁移和侵袭。CAFs可能通过外泌体将miR-889-3p转移至M0巨噬细胞。STAT1是miR-889-3p的靶标。此外，体内研究证实，源自CAFs的外泌体miR-889-3p可通过调节STAT1来加速ESCC肿瘤生长。源自CAFs的外泌体miR-889-3p通过下调STAT1抑制M1巨噬细胞极化，从而促进食管鳞状细胞癌细胞的增殖、迁移和侵袭，为ESCC提供了一个有前景的治疗靶点。
