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Neuropilin-2 acts a critical determinant for epithelial-to-mesenchymal transition and aggressive behaviors of human head and neck cancer

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Abstract

Purpose

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.

Methods

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.

Results

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.

Conclusions

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.

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Data Availability

All publicly accessed data are available on databases described in methodology. Analyzed datasets used during the study can be made available from the corresponding author upon reasonable request.

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Acknowledgements

This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIT) (2019R1A2C1085896 and 2020R1C1C1005480).

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Authors and Affiliations

  1. Department of Oral Pathology, School of Dentistry and Dental Research Institute, Seoul National University, Seoul, 03080, Republic of Korea

    Min-Hye Ahn, Ji-Hoon Kim, Su-Jung Choi, Hyun-Ji Kim, Dong-Guk Park, Hye-Jung Yoon, Seong-Doo Hong, Jae-Il Lee, Ji-Ae Shin & Sung-Dae Cho

  2. Chemical Biology Research Center, Korea Research Institute of Bioscience and Biotechnology, Ochang, Cheongwon, 28116, Republic of Korea

    Min-Hye Ahn

  3. Department of Oral Pathology, College of Dentistry, Dankook University, Cheonan, Republic of Korea

    Kyu-Young Oh

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  1. Min-Hye Ahn
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Contributions

M.-H.A. contributed to the conception, design, data acquisition, and analysis, and authored the original manuscript; J.-H.K., S.-J.C., H.-J.K., and D.-G.P. contributed to data acquisition and analyzed the in silico data; K.-Y.O., H.-J.Y., S.-D.H., and J.-I.L. participated in data analysis of the clinical samples; J.-A.S. and S.-D.C. contributed to the conception and design of experiments and supervised and authored the final manuscript. All authors approved the final manuscript and agreed to be responsible for all aspects of this work.

Corresponding authors

Correspondence to Ji-Ae Shin or Sung-Dae Cho.

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All procedures for clinical samples were performed after obtaining approval from the Institutional Review Board of the Seoul National University Dental Hospital (IRB number: ERI20021).

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Ahn, MH., Kim, JH., Choi, SJ. et al. Neuropilin-2 acts a critical determinant for epithelial-to-mesenchymal transition and aggressive behaviors of human head and neck cancer. Cell Oncol. 47, 497–511 (2024). https://doi.org/10.1007/s13402-023-00878-7

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