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Neuropathiazol induces neuronal-like differentiation in neuroblastoma cells via upregulation of PEG5

Pediatric Research volume 98, pages 1570–1579 (2025)Cite this article

Abstract

Background

Differentiation therapy is emerging as a promising strategy for treating neuroblastoma. However, the effects of neuropathiazol, a small molecule known to induce neuronal differentiation, have not been explored in neuroblastoma.

Procedure

Neuroblastoma cell lines were used to investigate the effects of neuropathiazol and retinoic acid on cell morphology, proliferation, and invasion in vitro. In vivo, neuroblastoma cells were implanted in nude mice to assess neuropathiazol’s therapeutic potential. Silver staining and markers of mature neurons were employed to evaluate neuropathiazol’s ability to promote neuronal differentiation.

Results

Neuropathiazol significantly inhibited the proliferation and invasion of neuroblastoma cells in vitro. It also enhanced synaptic growth and increased the expression of mature neuron markers more effectively than retinoic acid. Neuropathiazol treatment upregulated PEG5 expression, suggesting its role in promoting neuronal differentiation. Silencing PEG5 reversed these differentiation effects, reducing neuronal features. In vivo, neuropathiazol suppressed tumor growth and induced neuron-like differentiation in tumor tissues. However, its efficacy was diminished when PEG5 was knocked down. Additionally, neuropathiazol synergized with cyclophosphamide, enhancing its anti-neuroblastoma effects.

Conclusion

Neuropathiazol induces neuroblastoma differentiation, partly through PEG5 upregulation. As a promising differentiating agent for neuroblastoma, the combination of neuropathiazol and cyclophosphamide offers a potential treatment strategy for the disease.

Impact

  • Neuropathiazol significantly inhibits neuroblastoma cell proliferation and invasion in vitro.

  • Neuropathiazol promotes synaptic growth and upregulates mature neuronal marker expression more effectively than retinoic acid.

  • Neuropathiazol induces significant neuronal-like differentiation of neuroblastoma cells in vivo, leading to tumor growth suppression.

  • PEG5 is identified as a critical mediator of neuropathiazol’s differentiation-inducing effects. Knockdown of PEG5 reverses these effects, underscoring its pivotal role.

  • The combination of neuropathiazol with cyclophosphamide synergistically enhances anti-neuroblastoma effects, offering a compelling pharmacotherapeutic strategy.

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Fig. 1: Effects of RA and NPZ on inducing differentiation of neuroblastoma cells.
Fig. 2: Effects of RA and NPZ on the expression of MAP2, NEFL, and PEG5 proteins in neuroblastoma cells.
Fig. 3: Effect of PEG5 knockdown on neuroblastoma cell differentiation.
Fig. 4: Effects of NPZ and CTX treatments on SK-N-AS tumor-bearing nude mice.
Fig. 5: Effect of NPZ treatment on cellular differentiation in tumor tissues of SK-N-AS xenograft mice.

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Data availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Funding

This work was supported by the Jiangxi Provincial Health Commission Technology Plan Project (SKJP220219919).

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Author notes

  1. These authors contributed equally: Hao Xu, Fei Zhang.

Authors and Affiliations

  1. Department of Pediatrics, Wuhan NO.1 Hospital, Wuhan, 430022, China

    Hao Xu

  2. Department of Pediatrics, Wuhan Yaxin General Hospital, Wuhan, 430056, China

    Fei Zhang

  3. Department of Endoscopy Center, Jiangxi Cancer Hospital, The Second Affiliated Hospital of Nanchang Medical College, Jiangxi Clinical Research Center for Cancer, Nanchang, 330029, China

    Yi Xu

  4. Department of Pharmacy, Fengcheng People’s Hospital, Fengcheng, 331104, China

    Tianpeng Chen

  5. Department of Head and Neck Surgery, Jiangxi Cancer Hospital, The Second Affiliated Hospital of Nanchang Medical College, Nanchang, 330029, Jiangxi, China

    Fenqian Yuan

  6. Department of Oncology, Ganzhou People’s Hospital, Ganzhou, 341000, China

    Qihong Nie

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Contributions

Hao Xu: conceptualization, investigation, methodology, project administration, visualization, writing – original draft; Fei Zhang: data curation, formal analysis, methodology, software, writing – review & editing; Yi Xu: data curation, methodology, software, validation; Tianpeng Chen: methodology, resources; Fenqian Yuan: data curation, software, validation; Qihong Nie: funding acquisition, supervision.

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Correspondence to Qihong Nie.

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Xu, H., Zhang, F., Xu, Y. et al. Neuropathiazol induces neuronal-like differentiation in neuroblastoma cells via upregulation of PEG5. Pediatr Res 98, 1570–1579 (2025). https://doi.org/10.1038/s41390-025-03925-1

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