Cinobufagin injection induces gastric cancer cell ferroptosis by regulating the ACSL4-mediated lipid peroxidation pathway.

[UNLABELLED] This study investigates the anti-tumor mechanism of Cinobufagin (CBG) injection against gastric cancer. , CBG injection significantly suppressed xenograft tumor growth in nude mice. , it inhibited gastric cancer cell proliferation in a concentration-dependent manner and induced ferroptosis, evidenced by characteristic mitochondrial shrinkage, lipid reactive oxygen species accumulation, and an altered redox balance (e.g., elevated MDA, depleted GSH). To elucidate the underlying mechanism, we performed metabolomic profiling and Western blotting, which identified acyl-CoA synthetase long-chain family member 4 (ACSL4), a key regulator of lipid metabolism, as a critical target. CBG injection markedly upregulated ACSL4 expression. The functional relevance of ACSL4 was confirmed by rescue experiments: both pharmacological inhibition and genetic silencing of ACSL4 effectively reversed CBG injection-induced cell death and lipid peroxidation. In conclusion, CBG injection exerts its potent anti-gastric cancer effect by upregulating ACSL4. This upregulation drives lipid metabolic reprogramming, leading to lethal lipid peroxidation and subsequent ferroptosis. Our findings establish CBG injection as a promising therapeutic candidate for gastric cancer by targeting the ACSL4-dependent ferroptosis pathway.

[SUPPLEMENTARY INFORMATION] The online version contains supplementary material available at 10.1007/s10616-026-00937-5.