Periplocin Kills Osteosarcoma Cells by Driving Necroptosis and Pyroptosis

A new study published in Biology Direct reports that periplocin, a cardiotonic glycoside derived from traditional Chinese medicine, suppresses the progression of osteosarcoma, an aggressive bone tumor with poor prognosis, by triggering two forms of regulated cell death, necroptosis and pyroptosis, through the 4EBP1-eIF4E pathway. The peer-reviewed article, published open access on 16 October 2026, combines experiments in human osteosarcoma cell lines with a xenograft mouse model to map the mechanism.

The research team, led by Jianwei Sun, Shenghong Xu, Qi Liu and senior author Li Ding, treated human osteosarcoma and U2 OS cell lines with periplocin and measured cell viability, migration, proliferation and cell death using Cell Counting Kit-8, transwell, wound healing, colony formation and flow cytometry assays. Periplocin significantly inhibited osteosarcoma cell proliferation, migration and colony formation in a dose- and time-dependent manner, while markedly increasing cell death.

To determine which forms of cell death were responsible, the investigators examined key molecular markers of necroptosis and pyroptosis by Western blot. Periplocin treatment increased the levels of phosphorylated receptor-interacting protein kinase 3 (p-RIP3) and phosphorylated mixed lineage kinase domain-like protein (p-MLKL), hallmarks of necroptosis, as well as ASC, caspase-1, gasdermin D, interleukin-1 and NLRP3, markers associated with pyroptosis. When the cells were treated with necrostatin-1 (Nec-1), a necroptosis inhibitor, or disulfiram (DSF), a pyroptosis-related inhibitor, the effects of periplocin were attenuated, confirming that both death pathways contribute to the drug’s anticancer activity.

The study then probed the upstream regulatory mechanism centered on the 4EBP1-eIF4E pathway. Using several complementary approaches, eIF4E siRNA knockdown, pharmacological inhibition with 4EGI-1, overexpression of a phosphomimetic mutant eIF4E, and 4EBP1 overexpression, the researchers showed that eIF4E knockdown and inhibition, as well as 4EBP1 overexpression, reduced the osteosarcoma cell death induced by periplocin and promoted cell proliferation, migration and colony formation. Overexpression of the phosphomimetic mutant eIF4E partially reversed the tumor-promoting effects of eIF4E knockdown, pointing to the phosphorylation state of eIF4E as a critical determinant.

Co-immunoprecipitation experiments provided direct evidence for the interaction at the protein level: the binding between 4EBP1 and eIF4E significantly decreased after periplocin treatment. According to the authors’ conclusion, periplocin suppresses osteosarcoma progression through downregulation of 4EBP1, resulting in eIF4E activation and the subsequent induction of necroptosis and pyroptosis.

The antitumor effects were also validated in vivo. In a xenograft mouse model, periplocin suppressed tumor growth and enhanced necroptotic and pyroptotic markers in tumor tissues. When eIF4E was pharmacologically inhibited in this setting, the antitumor effects of periplocin were partially abolished, consistent with the cell culture findings and supporting the pathway’s relevance in living tumors.

Osteosarcoma is described in the study’s background as an aggressive bone tumor with poor prognosis, a situation the authors say highlights the need for novel therapeutic strategies. Periplocin has previously shown antitumor effects in various cancers, but its role in osteosarcoma had remained unclear before this work. The new findings, the authors write, highlight the therapeutic potential of periplocin and the 4EBP1-eIF4E pathway as promising targets for osteosarcoma treatment.

It is worth noting the scope and limitations of the evidence. The cell line work was performed in human osteosarcoma and U2 OS cell lines, and the in vivo validation used a xenograft mouse model rather than human patients. The authors note that the article is being shared early as a peer-reviewed, accepted, citable version carrying a permanent DOI, subject to further edits before the final Version of Record. The mechanistic claims rest on inhibitor-based experiments, siRNA knockdown, overexpression and co-immunoprecipitation within these preclinical systems.

Within those boundaries, the study offers a coherent mechanistic account: a natural-product glycoside disrupts the 4EBP1-eIF4E interaction, activating eIF4E signaling that culminates in necroptotic and pyroptotic death of osteosarcoma cells, with corresponding suppression of tumor growth in mice. Whether this translates into clinical benefit for osteosarcoma patients will require further research beyond the present preclinical models.

Subject of Research: Periplocin-induced necroptosis and pyroptosis in osteosarcoma through the 4EBP1-eIF4E pathway

Article Title: Periplocin suppresses osteosarcoma progression by triggering necroptosis and pyroptosis through the “4EBP1-eIF4E” pathway

Article References: Sun, J., Xu, S., Liu, Q., Yang, J., Yang, Y., Hao, S., Cheng, Y., & Ding, L. (2026). Periplocin suppresses osteosarcoma progression by triggering necroptosis and pyroptosis through the “4EBP1-eIF4E” pathway. Biology Direct. https://doi.org/10.1186/s13062-026-01021-w

Image Credits: AI Generated

DOI: 10.1186/s13062-026-01021-w

Keywords: periplocin, osteosarcoma, necroptosis, pyroptosis, 4EBP1, eIF4E, natural product, cell death, xenograft model, traditional Chinese medicine, Biology Direct, suppresses