Heydari Sheikhhossein Hamid, Amato Luisa, De Rosa Viviana, De Rosa Caterina, Ariano Annalisa, Critelli Sabrina, Omodei Daniela, Nele Valeria, Tuccillo Concetta, Franco Paola, Roviello Giovanni N, Camerlingo Rosa, Piattelli Adriano, Vicidomini Giovanni, Morgillo Floriana, De Rosa Giuseppe, Stoppelli Maria Patrizia, Della Corte Carminia Maria, Di Pietro Natalia, Iommelli Francesca
Department of Medical, Oral and Biotechnological Sciences, University "G. d'Annunzio" of Chieti-Pescara, 66100 Chieti, Italy.
Department of Precision Medicine, University of Campania Luigi Vanvitelli, 80131 Naples, Italy.
Int J Mol Sci. 2025 Jul 16;26(14):6825. doi: 10.3390/ijms26146825.
Increased IL-1β levels may promote carcinogenesis and metastasis by affecting tumor biology and the tumor microenvironment (TME). In this context, extracellular vesicles (EVs) play a key role in cell-to-cell communication, thus modulating the TME and immune response. Here, we aimed to test whether tumor-derived small EVs (TEVs) isolated from sensitive and osimertinib-resistant (OR) non-small-cell lung cancer (NSCLC) cells may promote EMT via fibronectin binding to α5β1 integrin as well as suppress the immune system and if these effects may be favored by IL-1β.
TEVs were isolated from control, OR, and IL-1β-stimulated NSCLC cells. Expressions of fibronectin and PD-L1 were screened in TEVs and the mRNA levels of vimentin and SMAD3 were also assessed in cancer cells after TEV co-culturing. Furthermore, to detect the effect on immune cells, we co-cultured TEVs with lung cancer patients' peripheral blood mononuclear cells (PBMCs).
TEVs were positive for fibronectin and the highest protein levels were found in TEVs obtained from the OR and IL-1β-stimulated cells. TEV-mediated activation of α5β1 signaling led to the upregulation of vimentin and SMAD3 mRNA in NSCLC cells and stimulated cell migration. EVs also increased PD-1, CTLA-4, FOXP3, TNF-α, IL-12, and INF-γ mRNA in lung cancer patients' immune cells.
Our findings indicate that TEVs promote EMT in NSCLC cells by the activation of the fibronectin-α5β1 axis. Finally, IL-1β stimulation induces TEV release with biological properties similar to OR TEVs, thus leading to cancer invasion and immune suppression and suggesting that inflammation can promote tumor spreading.
白细胞介素-1β（IL-1β）水平升高可能通过影响肿瘤生物学特性和肿瘤微环境（TME）促进肿瘤发生和转移。在此背景下，细胞外囊泡（EVs）在细胞间通讯中起关键作用，从而调节TME和免疫反应。在这里，我们旨在测试从敏感和奥希替尼耐药（OR）的非小细胞肺癌（NSCLC）细胞中分离出的肿瘤源性小细胞外囊泡（TEVs）是否可通过纤连蛋白与α5β1整合素结合促进上皮-间质转化（EMT），以及是否会抑制免疫系统，以及IL-1β是否会促进这些作用。
从对照、OR和IL-1β刺激的NSCLC细胞中分离TEVs。在TEVs中筛选纤连蛋白和程序性死亡配体1（PD-L1）的表达，并在TEV共培养后评估癌细胞中波形蛋白和SMAD3的mRNA水平。此外，为了检测对免疫细胞的影响，我们将TEVs与肺癌患者的外周血单个核细胞（PBMCs）共培养。
TEVs纤连蛋白呈阳性，在从OR和IL-1β刺激的细胞中获得的TEVs中发现最高蛋白水平。TEV介导的α5β1信号激活导致NSCLC细胞中波形蛋白和SMAD3 mRNA上调并刺激细胞迁移。EVs还增加了肺癌患者免疫细胞中程序性死亡蛋白1（PD-1）、细胞毒性T淋巴细胞相关蛋白4（CTLA-4）、叉头框蛋白P3（FOXP3）、肿瘤坏死因子-α（TNF-α）、白细胞介素-12（IL-12）和干扰素-γ（INF-γ）的mRNA水平。
我们的研究结果表明，TEVs通过激活纤连蛋白-α5β1轴促进NSCLC细胞的EMT。最后，IL-1β刺激诱导具有与OR TEVs相似生物学特性的TEV释放，从而导致癌症侵袭和免疫抑制，并表明炎症可促进肿瘤扩散。
