Bourreau Clara, Navarro Emilie, Cotinat Marine, Krejbich Morgane, Guillonneau François, Guette Catherine, Boissard Alice, Henry Cécile, Corre Isabelle, Treps Lucas, Clere Nicolas
Univ Angers, Inserm, CNRS, MINT, SFR ICAT, Angers, France.
Nantes Université, Université d'Angers, CHU Nantes, Inserm, CNRS, CRCI2NA, Nantes, France.
Cancer Med. 2025 Mar;14(5):e70707. doi: 10.1002/cam4.70707.
The tumor microenvironment (TME) of non-small cell lung cancer (NSCLC) is highly heterogeneous and is involved in tumorigenesis and resistance to therapy. Among the cells of the TME, endothelial cells are associated with the latter processes through endothelial-to-mesenchymal transition (EndMT). During EndMT, endothelial cells (ECs) progressively lose their endothelial phenotype in favor of a mesenchymal phenotype, which favors the production of cancer-associated fibroblasts (CAFs). Our study aimed to investigate the consequences of exposure to different lung tumor secretomes on EC phenotype and plasticity.
Conditioned media (CM) were prepared from the tumor cell lines A549, H1755, H23, H1437, and H1975. Proliferation and migration of ECs treated with these CMs were assessed by Cyquant and Incucyte technologies, respectively. The angiogenic capacity of ECs was assessed by following tubulogenesis on Matrigel. Phenotypic changes in treated ECs were detected by flow cytometry. Morphological analysis of actin fibers was performed by immunohistochemistry, while proteomic analysis by mass spectrometry was used to identify the protein content of secretomes.
A change of the endothelial phenotype was found when human umbilical vein endothelial cells (HUVECs) were treated with different CMs. This phenotypic change was associated with a morphological change, an increase in both stress fiber expression and spontaneous migration. Furthermore, an increase in mesenchymal markers (α-SMA and CD44) confirmed the phenotypic changes. However, the secretomes did not modify the rate of double-labeled cells (vWF/α-SMA or CD31/CD44). Proteomic analysis identified potential targets involved in the EndMT with therapeutic relevance.
Taken together, these data suggest that CMs can induce partial EndMT.
非小细胞肺癌（NSCLC）的肿瘤微环境（TME）高度异质性，参与肿瘤发生和治疗抵抗。在TME的细胞中，内皮细胞通过内皮-间充质转化（EndMT）与后一过程相关。在EndMT过程中，内皮细胞（ECs）逐渐失去其内皮表型，转而形成间充质表型，这有利于癌相关成纤维细胞（CAFs）的产生。我们的研究旨在探讨暴露于不同肺肿瘤分泌组对EC表型和可塑性的影响。
从肿瘤细胞系A549、H1755、H23、H1437和H1975制备条件培养基（CM）。分别通过Cyquant和Incucyte技术评估用这些CM处理的ECs的增殖和迁移。通过在基质胶上观察管形成来评估ECs的血管生成能力。通过流式细胞术检测处理后ECs的表型变化。通过免疫组织化学对肌动蛋白纤维进行形态学分析，同时使用质谱进行蛋白质组分析以鉴定分泌组的蛋白质含量。
当人脐静脉内皮细胞（HUVECs）用不同的CM处理时，发现内皮表型发生了变化。这种表型变化与形态学变化、应激纤维表达增加和自发迁移增加有关。此外，间充质标志物（α-SMA和CD44）的增加证实了表型变化。然而，分泌组并未改变双标记细胞（vWF/α-SMA或CD31/CD44）的比例。蛋白质组分析确定了与EndMT相关且具有治疗相关性的潜在靶点。
综上所述，这些数据表明CM可以诱导部分EndMT。
