Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, 4710-057, Braga, Portugal.
ICVS/3B's-PT Government Associate Laboratory, Braga/Guimarães, Portugal.
Cell Oncol (Dordr). 2020 Aug;43(4):643-654. doi: 10.1007/s13402-020-00510-y. Epub 2020 Mar 29.
Previously, inflammation has been found to be associated with the development of lung cancer. Despite their well-characterized pro-inflammatory functions, the putative roles of interleukin-17 (IL-17) cytokine family members in tumorigenesis have remained controversial. While IL-17A exhibits both pro- and anti-tumor effects, IL-17F has been suggested to serve as a candidate for cancer therapy. Thus, we aimed at clarifying the involvement of IL-17A/F in lung cancer.
IL-17 receptor expression in human and murine lung cancer cells was assessed using immunofluorescence. The effect of IL-17A/F stimulation on lung cancer cell viability (SRB assay) and metabolism (glucose consumption and lactate production) was evaluated under normoxic and hypoxic conditions. Characterization of IL-17A/F-stimulated macrophages was performed by flow cytometry and ELISA. The effect of conditioned media (CM) from IL-17A/F-stimulated macrophages was evaluated on lung cancer cell migration. The effect of CM-stimulated macrophages on lung tumor growth, proliferation and angiogenesis was evaluated in vivo using a chicken chorioallantoic membrane (CAM) assay.
No alterations in lung cancer cell viability or metabolism were observed upon direct stimulation with IL-17A/F. We found, however, that CM from IL-17A/F-stimulated macrophages promoted both murine and human lung cancer cell progression through an increased migration capacity in vitro and enhanced in vivo tumor growth, proliferation and angiogenesis. These findings were supported by an increased polarization of human macrophages towards a M2-like phenotype.
Our data indicate that IL-17A/F act through immune cell orchestration, i.e., of macrophages, to promote lung cancer cell growth and progression. In addition, our data provide a link between IL-17A/F activity and lung cancer cell-macrophage crosstalk.
先前发现炎症与肺癌的发展有关。尽管白细胞介素-17（IL-17）细胞因子家族成员具有明确的促炎功能，但它们在肿瘤发生中的潜在作用仍存在争议。虽然 IL-17A 表现出促肿瘤和抗肿瘤作用，但 IL-17F 被认为是癌症治疗的候选药物。因此，我们旨在阐明 IL-17A/F 在肺癌中的作用。
使用免疫荧光法评估人源和鼠源肺癌细胞中 IL-17 受体的表达。在常氧和低氧条件下，通过 SRB 测定法评估 IL-17A/F 刺激对肺癌细胞活力的影响，通过葡萄糖消耗和乳酸产生评估代谢的影响。通过流式细胞术和 ELISA 对 IL-17A/F 刺激的巨噬细胞进行特征分析。通过评价 IL-17A/F 刺激的巨噬细胞条件培养基（CM）对肺癌细胞迁移的影响来评估其作用。通过鸡胚绒毛尿囊膜（CAM）试验评估 CM 刺激的巨噬细胞对体内肺肿瘤生长、增殖和血管生成的影响。
直接用 IL-17A/F 刺激未观察到肺癌细胞活力或代谢的改变。然而，我们发现 IL-17A/F 刺激的巨噬细胞 CM 通过体外增强的迁移能力和体内增强的肿瘤生长、增殖和血管生成促进了人和鼠源肺癌细胞的进展。这些发现得到了人源巨噬细胞向 M2 样表型极化增加的支持。
我们的数据表明，IL-17A/F 通过免疫细胞的协调作用，即巨噬细胞，促进肺癌细胞的生长和进展。此外，我们的数据提供了 IL-17A/F 活性与肺癌细胞-巨噬细胞相互作用之间的联系。
