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Background Non-small cell lung cancer (NSCLC) remains a leading cause of cancer-related mortality globally. Although chemotherapy and targeted therapies provide clinical benefits, their efficacy is often limited by drug resistance and the immunosuppressive tumor microenvironment. 1–3 Recently, plant-derived saponins have attracted growing interest for their ability to stimulate antitumor immune responses by inducing immunogenic cell death (ICD), a form of cell death characterized by the release of damage-associated molecular patterns (DAMPs), such as HMGB1 and ATP.4 Akebia quinata, a saponin-rich medicinal plant, has shown various pharmacological properties. However, its immunomodulatory potential in NSCLC remains largely unexplored. This study aims to investigate the anticancer effects of Akebia quinata saponins (AQS) with a focus on their ability to induce apoptosis and ICD in NSCLC cells.Methods AQS were extracted from dried seeds of A. quinata using 70% ethanol and concentrated under reduced pressure. Their chemical constituents were qualitatively profiled and tentatively identified using UHPLC-QTOF MS/MS. Human NSCLC cell lines A549 and H460 were treated with various concentrations of AQS to evaluate their anticancer effects. Apoptosis was assessed by Annexin V staining and sub-G1 population analysis. Western blotting was performed to investigate the signaling pathway regulated by AQS. To investigate ICD, extracellular HMGB1 levels were measured using ELISA, and ATP release was quantified using a luminescence-based assay.Results AQS treatment resulted in a dose-dependent decrease in cell viability, indicating potent cytotoxic effects. Flow cytometric analysis revealed a significant increase in Annexin V-positive cells, along with an elevated sub-G1 population, confirming the induction of apoptosis. These findings were further supported by increased expression of cleaved caspases and PARP1 in AQS-treated cells. Mechanistically, AQS treatment led to notable changes in the MAPK signaling pathway. In addition to apoptotic effects, AQS demonstrated the ability to trigger ICD. Measurement of extracellular DAMPs revealed that AQS significantly increased the release of HMGB1 and ATP into the culture supernatant. Collectively, these results suggest that AQS not only induces cancer cell apoptosis but also promotes immunogenic features that may enhance anticancer immunity.Conclusions These findings indicate that AQS exerts anticancer effects in NSCLC cells by inducing apoptosis and ICD, potentially through the MAPK signaling pathway. AQS may serve as a promising candidate for the development of novel therapeutic strategies against NSCLC. Further studies are warranted to fully explore the therapeutic potential of AQS as an anticancer agent.References Zhu L, Wang Y, Lv W, et al. Schizandrin A can inhibit non-small cell lung cancer cell proliferation by inducing cell cycle arrest, apoptosis and autophagy. Int J Mol Med. 2021;48:214.Duma N, Santana-Davila R, Molina JR. Non-small cell lung cancer: epidemiology, screening, diagnosis, and treatment. Mayo Clin Proc. 2019;94:1623–1640.Alexander M, Kim SY, Cheng H. Update 2020: management of non-small cell lung cancer. Lung. 2020;198:897–907.Dong S, Guo X, Han F, et al. Emerging role of natural products in cancer immunotherapy. Acta Pharm Sin B. 2022;12:1163–1185.