Fang Hua, Chi Xiaowen, Wang Mengyao, Liu Jing, Sun Meiqi, Zhang Jiashu, Zhang Wei
Respiratory and Critical Care Medicine, The First Affiliated Hospital of Harbin Medical University, No. 23, Post Street, Nangang District, Harbin, 150001, China.
Respiratory and Critical Care Medicine, The Second Hospital of Heilongjiang Province, Harbin, 150028, China.
Mol Cell Biochem. 2025 May;480(5):3019-3032. doi: 10.1007/s11010-024-05161-3. Epub 2024 Nov 29.
Tumor-associated macrophages (TAMs) are a type of highly plastic immune cells in the tumor microenvironment (TME), which can be classified into two main phenotypes: classical activated M1 macrophages and alternatively activated M2 macrophages. As previously reported, M2-polarized TAMs play critical role in promoting the progression of non-small cell lung cancer (NSCLC) via secreting exosomes, but the detailed mechanisms are still largely unknown. In the present study, the THP-1 monocytes were sequentially induced into M0 and M2-polarized macrophages, and the exosomes were obtained from M0 (M0-exos) and M2 (M2-exos) polarized macrophages, respectively, and co-cultured with NSCLC cells (H1299 and A549) to establish the exosomes-cell co-culture system in vitro. As it was determined by MTT assay, RT-qPCR and Transwell assay, in contrast with the M0-exos, M2-exos significantly promoted cell proliferation, migration and epithelial-mesenchymal transition (EMT) process in NSCLC cells. Next, through screening the contents in the exosomes, it was verified that miR-155-5p was especially enriched in the M2-exos, and M2-exos enhanced cancer aggressiveness and tumorigenesis in in vitro NSCLC cells and in vivo xenograft tumor-bearing mice models via delivering miR-155-5p. The detailed molecular mechanisms were subsequently elucidated, and it was found that miR-155-5p bound with HuR to increase the stability and expression levels of VEGFR2, which further activated the tumor-promoting PI3K/Akt/mTOR signal pathway, and M2-exos-enhanced cancer progression in NSCLC cells were apparently suppressed by downregulating VEGFR2 and PI3K inhibitor LY294002 co-treatment. Taken together, M2-polarized TAMs secreted miR-155-5p-containing exosomes to enhanced cancer aggressiveness of NSCLC by activating the VEGFR2/PI3K/Akt/mTOR pathway in a HuR-dependent manner.
肿瘤相关巨噬细胞(TAMs)是肿瘤微环境(TME)中一类高度可塑性的免疫细胞,可分为两种主要表型:经典活化的M1巨噬细胞和替代性活化的M2巨噬细胞。如先前报道,M2极化的TAMs通过分泌外泌体在促进非小细胞肺癌(NSCLC)进展中起关键作用,但其详细机制仍大多未知。在本研究中,将THP-1单核细胞依次诱导为M0和M2极化巨噬细胞,分别从M0(M0-exos)和M2(M2-exos)极化巨噬细胞中获得外泌体,并与NSCLC细胞(H1299和A549)共培养以在体外建立外泌体-细胞共培养系统。通过MTT法、RT-qPCR和Transwell法测定,与M0-exos相比,M2-exos显著促进NSCLC细胞的增殖、迁移和上皮-间质转化(EMT)过程。接下来,通过筛选外泌体中的成分,证实miR-155-5p在M2-exos中特别富集,并且M2-exos通过递送miR-155-5p在体外NSCLC细胞和体内异种移植荷瘤小鼠模型中增强癌症侵袭性和肿瘤发生。随后阐明了详细的分子机制,发现miR-155-5p与HuR结合以增加VEGFR2的稳定性和表达水平,这进一步激活了促肿瘤的PI3K/Akt/mTOR信号通路,并且通过下调VEGFR2和PI3K抑制剂LY294002联合处理明显抑制了M2-exos增强的NSCLC细胞中的癌症进展。综上所述,M2极化的TAMs分泌含miR-155-5p的外泌体,以HuR依赖的方式激活VEGFR2/PI3K/Akt/mTOR途径,从而增强NSCLC的癌症侵袭性。