Department of Oral Pathology, School of Dentistry and Dental Research Institute, Seoul National University, Seoul, 03080, Republic of Korea.
Chemical Biology Research Center, Korea Research Institute of Bioscience and Biotechnology, Ochang, Cheongwon, 28116, Republic of Korea.
Cell Oncol (Dordr). 2024 Apr;47(2):497-511. doi: 10.1007/s13402-023-00878-7. Epub 2023 Oct 3.
Neuropilin-2 (NRP2) is a multifunctional single-pass transmembrane receptor that binds to two disparate ligands, namely, vascular endothelial growth factors (VEGFs) and semaphorins (SEMAs). It is reportedly involved in neuronal and vascular development. In this study, we uncovered the exact functional role of NRP2 and its molecular mechanism during aggressive behaviors and lymph node (LN) metastasis in human head and neck cancer (HNC) and identified algal methanol extract as a potential novel NRP2 inhibitor.
In silico analyses and immunohistochemistry were used to investigate the relationship between NRP2 expression and the prognosis of HNC patients. The functional role of NRP2 on the proliferation, migration, invasion, and cancer stem cell (CSC) properties of HNC cells was examined by MTS, soft agar, clonogenic, transwell migration and invasion assays, and sphere formation assays. Signaling explorer antibody array, western blot, and qPCR were performed toward the investigation of a molecular mechanism that is related to NRP2.
NRP2 was highly expressed in HNC and positively correlated with LN metastasis and advanced tumor stage and size in patients. Using loss- or gain-of-function approaches, we found that NRP2 promoted the proliferative, migratory, and invasive capacities of human HNC cells. Furthermore, NRP2 regulated Sox2 expression to exhibit aggressiveness and CSC properties of human HNC cells. We demonstrated that p90 ribosomal S6 kinase 1 (RSK1) elevates the aggressiveness and CSC properties of human HNC cells, possibly by mediating NRP2 and Sox2. Zeb1 was necessary for executing the NRP2/RSK1/Sox2 signaling pathway during the induction of epithelial-to-mesenchymal transition (EMT) and aggressive behaviors of human HNC cells. Moreover, the methanol extract of Codium fragile (MECF) repressed NRP2 expression, inhibiting the RSK1/Sox2/Zeb1 axis, which contributed to the reduction of aggressive behaviors of human HNC cells.
These findings suggest that NRP2 is a critical determinant in provoking EMT and aggressive behaviors in human HNC through the RSK1/Sox2/Zeb1 axis, and MECF may have the potential to be a novel NRP2 inhibitor for treating metastasis in HNC patients.
神经纤毛蛋白 2（NRP2）是一种多功能单次跨膜受体，可与两种截然不同的配体结合，即血管内皮生长因子（VEGFs）和神经鞘氨醇（SEMA）。据报道，它参与神经元和血管的发育。在这项研究中，我们揭示了 NRP2 在人类头颈部癌症（HNC）侵袭性行为和淋巴结（LN）转移中的确切功能作用及其分子机制，并确定藻类甲醇提取物是一种潜在的新型 NRP2 抑制剂。
通过计算机分析和免疫组织化学分析，研究了 NRP2 表达与 HNC 患者预后的关系。通过 MTS、软琼脂、集落形成、Transwell 迁移和侵袭实验以及球体形成实验，研究了 NRP2 对 HNC 细胞增殖、迁移、侵袭和癌症干细胞（CSC）特性的功能作用。通过信号转导探索抗体阵列、Western blot 和 qPCR 对与 NRP2 相关的分子机制进行了研究。
NRP2 在 HNC 中高表达，与 LN 转移以及患者的晚期肿瘤分期和大小呈正相关。通过失活或过表达的方法，我们发现 NRP2 促进了人 HNC 细胞的增殖、迁移和侵袭能力。此外，NRP2 调节 Sox2 的表达，从而表现出人 HNC 细胞的侵袭性和 CSC 特性。我们证明，p90 核糖体 S6 激酶 1（RSK1）通过介导 NRP2 和 Sox2 来提高人 HNC 细胞的侵袭性和 CSC 特性。Zeb1 是执行 NRP2/RSK1/Sox2 信号通路所必需的，该信号通路在诱导人 HNC 细胞上皮间质转化（EMT）和侵袭性行为中起作用。此外，Codium fragile 的甲醇提取物（MECF）抑制 NRP2 的表达，抑制 RSK1/Sox2/Zeb1 轴，从而减少人 HNC 细胞的侵袭性行为。
这些发现表明，NRP2 通过 RSK1/Sox2/Zeb1 轴在引发人 HNC 的 EMT 和侵袭性行为中是一个关键决定因素，而 MECF 可能具有作为治疗 HNC 患者转移的新型 NRP2 抑制剂的潜力。
