三阴性乳腺癌（TNBC）是一种异质性和侵袭性较强的癌症，具有高血管生成能力和频繁的转移。我们已经发现天然黄酮类化合物栓翼蛋白（SC）通过在体内规范肿瘤血管结构来抑制TNBC的自发性转移。在本研究中，肿瘤坏死因子α（TNFα）处理后的人乳腺微血管内皮细胞（HMMEC）培养上清液促进了TNBC细胞的迁移和伪足形成，但这些现象在与SC联合处理的HMMEC中消失。TNFα增强了HMMEC和人脐静脉内皮细胞（HUVEC）中粒细胞集落刺激因子（G-CSF）和粒细胞-巨噬细胞集落刺激因子（GM-CSF）的表达。G-CSF促进了TNBC的体外迁移和侵袭，而G-CSF中和抗体和SC均抑制了Balb/c小鼠中的TNBC转移。SC对G-CSF诱导的TNBC细胞迁移无抑制作用，但降低了TNBC肿瘤组织和TNFα刺激的内皮细胞（ECs）中的G-CSF含量。SC限制了RUNX1（跑读相关转录因子1）在TNBC肿瘤血管和TNFα处理的ECs中的核转位。发现RUNX1直接结合到TNBC肿瘤血管的G-CSF启动子上，并调控G-CSF的表达。TNF受体2（TNFR2）对调控TNFα诱导的RUNX1激活和G-CSF表达至关重要。值得注意的是，SC通过与TNFR2结合来阻断TNFα和TNFR2之间的相互作用。本研究证明了SC通过靶向TNFα/TNFR2引发的RUNX1激活和随后的G-CSF在TNBC相关ECs中的产生，从而降低了TNBC的转移。版权归Elsevier Inc.所有，保留所有权利。

Triple-negative breast cancer (TNBC) is heterogeneous and aggressive, with high vascularity and frequent metastasis. We have already found natural flavonoid scutellarin (SC) suppressed spontaneous TNBC metastasis via normalizing tumor vasculature in vivo. In this study, supernatant from tumor necrosis factorα (TNFα)-treated human mammary microvascular endothelial cell (HMMEC) promoted cell migration and pseudopod formation in TNBC cells, but these phenomena were disappeared in SC-co-treated HMMEC. TNFα enhanced the expression of granulocyte colony-stimulating factor (G-CSF) and granulocyte-macrophage colony-stimulating factor (GM-CSF) in both HMMEC and human umbilical vein endothelial cell (HUVEC). G-CSF promoted TNBC migration and invasion in vitro, while G-CSF neutralization antibody and SC both inhibited TNBC metastasis in Balb/c mice. SC had no inhibition on the G-CSF-induced TNBC cell migration, but reduced G-CSF content in TNBC tumor tissues and TNFα-stimulated endothelial cells (ECs). SC restricted the nuclear translocation of runt-related transcription factor 1 (RUNX1) in TNBC tumor vessels and TNFα-treated ECs. RUNX1 was found to directly bind to the promoter of G-CSF in TNBC tumor vessels and regulated G-CSF expression. TNF receptor 2 (TNFR2) was crucial for regulating the TNFα-induced RUNX1 activation and G-CSF expression. Notably, SC hindered the interaction between TNFα and TNFR2 via binding to TNFR2. This work demonstrated that SC reduced TNBC metastasis by targeting TNFα/TNFR2-initiated RUNX1 activation and subsequent G-CSF production in TNBC-associated ECs.Copyright © 2023 Elsevier Inc. All rights reserved.