Semaphorin 3F inhibits breast cancer metastasis by regulating the Akt-mTOR and TGFβ signaling pathways via neuropilin-2

Sci Rep. 2025 Mar 3;15(1):7394. doi: 10.1038/s41598-025-91559-y.

Abstract

Class 3 semaphorins are axon guidance factors implicated in tumor and vascular biology, including invasive activity. Recent studies indicate that semaphorin 3F (SEMA3F) is a potent inhibitor of metastasis; however, its functional role in breast cancer is not fully understood. We found that exogenous SEMA3F inhibited phosphorylation of Akt and mTOR downstream kinase S6K in MDA-MB-231 and MCF7 cells via neuropilin-2 (NRP2) receptor. We also examined the effect of SEMA3F on breast cancer progression in vivo allograft model. The mouse 4T1 breast cancer cells or 4T1 cells overexpressing SEMA3F (4T1-SEMA3F) were implanted into mammary fat pads of Balb/c mice. We found that tumor growth was significantly inhibited in 4T1-SEMA3F injected mice compared to controls. Immunostaining revealed a remarkable reduction in the expression of vimentin, a mesenchymal cell marker, in 4T1-SEMA3F tumors. We also observed that mice injected with 4T1-SEMA3F cells had minimal metastasis to the liver and lungs, compared to controls. As a novel feature, SEMA3F suppressed TGFβ-induced Smad2 phosphorylation, resulting in the inhibition of cell invasiveness and epithelial-to-mesenchymal transition (EMT) in breast cancer. Consistently, a significant correlation between reduced expression of SEMA3F and poor outcome in patients with breast cancer. We conclude that SEMA3F acts as a dual inhibitor of the Akt-mTOR and TGFβ signaling pathways; thus, it has the potential to treat metastatic breast cancer.

Keywords: Breast cancer; Metastasis; Neuropilin-2; Semaphorin 3F.

MeSH terms

  • Animals
  • Breast Neoplasms* / genetics
  • Breast Neoplasms* / metabolism
  • Breast Neoplasms* / pathology
  • Cell Line, Tumor
  • Female
  • Humans
  • MCF-7 Cells
  • Membrane Proteins* / genetics
  • Membrane Proteins* / metabolism
  • Mice
  • Mice, Inbred BALB C
  • Neoplasm Metastasis
  • Nerve Tissue Proteins* / genetics
  • Nerve Tissue Proteins* / metabolism
  • Neuropilin-2* / metabolism
  • Phosphorylation
  • Proto-Oncogene Proteins c-akt* / metabolism
  • Signal Transduction*
  • TOR Serine-Threonine Kinases* / metabolism
  • Transforming Growth Factor beta* / metabolism

Substances

  • Neuropilin-2
  • TOR Serine-Threonine Kinases
  • Proto-Oncogene Proteins c-akt
  • Transforming Growth Factor beta
  • Nerve Tissue Proteins
  • SEMA3F protein, human
  • Membrane Proteins
  • neuropilin-2, human
  • MTOR protein, human