Shogaol triggers apoptosis and ferroptosis in Ewing sarcoma by targeting the polo-like kinase 1/nucleophosmin 1 axis.

Ewing sarcoma is an aggressive pediatric bone cancer with poor prognosis, highlighting the urgent need for novel therapeutics. This study demonstrates the potent anti-tumor activity of shogaol, a bioactive compound from ginger, against Ewing sarcoma, using both in vitro and in vivo model systems. In vitro, shogaol significantly inhibited cell proliferation and induced both apoptosis and ferroptosis in Ewing sarcoma cells, as evidenced by increased lipid reactive oxygen species, Malondialdehyde, and Fe2+ levels, as well as altered expression of BAX, BCL2, GPX4, and SLC3A2. In vivo, shogaol significantly inhibited tumor cell growth in a subcutaneous xenograft mouse model using SKNMC cells. Using transcriptome sequencing and proteomics, we found that shogaol targeted the PLK1/NPM1 axis: it downregulated polo-like kinase 1 (PLK1), leading to reduced phosphorylation of NPM1 at Thr199. This facilitated the increased interaction between the E3 ligase TRIM28 and NPM1, leading to its ubiquitination and subsequent degradation, which in turn inhibited the MAPK signaling pathway. Key findings were validated by CETSA, isothermal dose-response fingerprinting, co-immunoprecipitation, and ubiquitination assays. These results demonstrate that shogaol exerts dual cytotoxic effects through the PLK1/NPM1 axis, presenting a potential natural therapeutic approach for Ewing sarcoma.