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Tumor suppressive functions of WNT5A in rhabdomyosarcoma
Nada Ragab1, Julia Bauer1, Anja Uhmann1
1Institute of Human Genetics, University Medical Center Göttingen, D‑37073 Göttingen, Germany.
Abstract:
Rhabdomyosarcoma (RMS) is a highly aggressive soft tissue malignancy that predominantly affects children. The main subtypes are alveolar RMS (ARMS) and embryonal RMS (ERMS) and the two show an impaired muscle differentiation phenotype. One pathway involved in muscle differentiation is WNT signaling. However, the role of this pathway in RMS is far from clear. Our recent data showed that the canonical WNT/β‑Catenin pathway serves a subordinate role in RMS, whereas non‑canonical WNT signaling probably is more important for this tumor entity. The present study investigated the role of WNT5A, which is the major ligand of non‑canonical WNT signaling, in ERMS and ARMS. Gene expression analysis showed that WNT5A was expressed in human RMS samples and that its expression is more pronounced in ERMS. When stably overexpressed in RMS cell lines, WNT5A decreased proliferation and migration of the cells as demonstrated by BrdU incorporation and Transwell migration or scratch assay, respectively. WNT5A also decreased the self‑renewal capacity and the expression of stem cell markers and modulates the levels of muscle differentiation markers as shown by sphere assay and western blot analysis, respectively. Finally, overexpression of WNT5A can destabilize active β‑Catenin of RMS cells. A WNT5A knockdown has opposite effects. Together, the results suggest that WNT5A has tumor suppressive functions in RMS, which accompanies downregulation of β‑Catenin.
Insights
WNT5A exhibits tumor-suppressive functions in rhabdomyosarcoma (RMS), a pediatric cancer. Overexpressing WNT5A reduces RMS cell proliferation, migration, and self-renewal, while downregulating beta-catenin.
Area of Science:
- Oncology
- Molecular Biology
- Cell Signaling
Background:
- Rhabdomyosarcoma (RMS) is an aggressive pediatric soft tissue cancer with impaired muscle differentiation.
- WNT signaling pathways are implicated in muscle differentiation, but their role in RMS remains unclear.
- Non-canonical WNT signaling, particularly WNT5A, is hypothesized to be more significant than canonical WNT/β-Catenin in RMS.
Purpose of the Study:
- To investigate the role of WNT5A, a key non-canonical WNT ligand, in embryonal RMS (ERMS) and alveolar RMS (ARMS).
- To determine the functional impact of WNT5A on RMS cell proliferation, migration, self-renewal, and differentiation markers.
Main Methods:
- Gene expression analysis of WNT5A in human RMS samples.
- Stable overexpression of WNT5A in RMS cell lines.
- Assays for cell proliferation (BrdU incorporation), migration (Transwell, scratch assay), and self-renewal (sphere assay).
- Western blot analysis to assess muscle differentiation markers and β-Catenin levels.
Main Results:
- WNT5A is expressed in human RMS, with higher levels in ERMS.
- WNT5A overexpression reduced RMS cell proliferation, migration, and self-renewal capacity.
- WNT5A modulated muscle differentiation markers and destabilized active β-Catenin.
- WNT5A knockdown produced opposite effects, indicating tumor-suppressive functions.
Conclusions:
- WNT5A acts as a tumor suppressor in RMS.
- WNT5A exerts its effects by reducing proliferation and migration and downregulating β-Catenin.
- These findings highlight WNT5A as a potential therapeutic target in RMS.
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