Sheng Youjing, Peng Weidong, Huang Yan, Cheng Lanqing, Meng Ye, Kwantwi Louis Boafo, Yang Jiezhen, Xu Jiegou, Xiao Han, Kzhyshkowska Julia, Wu Qiang
Department of Pathology, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Shushan District, Hefei 230022, Anhui, PR China.
Institute of Transfusion Medicine and Immunology, Mannheim Institute for Innate Immunoscience (MI3), Medical Faculty Mannheim, University of Heidelberg, Ludolf-Krehl Strasse 13-17, 68167 Mannheim, Germany.
J Leukoc Biol. 2023 Mar 29;113(4):383-399. doi: 10.1093/jleuko/qiad004.
The immune component of the tumor microenvironment is essential for the regulation of cancer progression. In breast cancer (BC), a patient's tumor mass is frequently infiltrated by neutrophils (tumor-associated neutrophils, TANs). Our study addressed the role of TANs and their mechanism of action in BC. Using quantitative IHC, ROC, and Cox analysis, we demonstrated that a high density of TANs infiltrating the tumor parenchyma was predictive of poor prognosis and of decreased progression-free survival of patients with BC, who underwent surgical tumor removal without previous neoadjuvant chemotherapy, in 3 different cohorts: training, validation, and independent cohorts. Conditioned medium from human BC cell lines prolonged the lifespan of healthy donor neutrophils ex vivo. Neutrophils activated by the supernatants of BC lines demonstrated an increased ability to stimulate proliferation, migration, and invasive activity of BC cells. Cytokines involved in this process were identified using antibody arrays. The relationship between these cytokines and the density of TANs was validated by ELISA and IHC in fresh BC surgical samples. It was determined that tumor-derived G-CSF significantly extended the lifespan and increased the metastasis-promoting activities of neutrophils via the PI3K-AKT and NF-κB pathways. Simultaneously, TAN-derived RLN2 promoted the migratory abilities of MCF7 cells via PI3K-AKT-MMP-9. Analysis of tumor tissues from 20 patients with BC identified a positive correlation between the density of TANs and the activation of the G-CSF-RLN2-MMP-9 axis. Finally, our data demonstrated that TANs in human BC have detrimental effects, supporting malignant cell invasion and migration.
肿瘤微环境的免疫成分对于癌症进展的调控至关重要。在乳腺癌（BC）中，患者的肿瘤块常常被中性粒细胞浸润（肿瘤相关中性粒细胞，TANs）。我们的研究探讨了TANs在BC中的作用及其作用机制。通过定量免疫组化、受试者工作特征曲线（ROC）分析和Cox分析，我们证明，在3个不同队列（训练队列、验证队列和独立队列）中，浸润肿瘤实质的高密度TANs可预测接受手术切除肿瘤且未接受过新辅助化疗的BC患者预后不良以及无进展生存期缩短。来自人BC细胞系的条件培养基可在体外延长健康供体中性粒细胞的寿命。被BC细胞系上清液激活的中性粒细胞刺激BC细胞增殖、迁移和侵袭活性的能力增强。使用抗体芯片鉴定了参与此过程的细胞因子。通过酶联免疫吸附测定（ELISA）和免疫组化在新鲜的BC手术样本中验证了这些细胞因子与TANs密度之间的关系。已确定肿瘤来源的粒细胞集落刺激因子（G-CSF）通过PI3K-AKT和NF-κB途径显著延长中性粒细胞的寿命并增强其促进转移的活性。同时，TAN来源的松弛素2（RLN2）通过PI3K-AKT-基质金属蛋白酶9（MMP-9）促进MCF7细胞的迁移能力。对20例BC患者的肿瘤组织分析发现，TANs密度与G-CSF-RLN2-MMP-9轴的激活之间呈正相关。最后，我们的数据表明，人BC中的TANs具有有害作用，支持恶性细胞的侵袭和迁移。
