Cauloside A interacts with TLR4 to induce JNK/Caspase-3/GSDME-dependent pyroptosis in non-small cell lung cancer cells

With non-small cell lung Cancer (NSCLC) accounts for the majority of diagnoses, lung Cancer continues to be the most common and deadly type of Cancer worldwide. While treatment options have advanced, therapeutic efficacy remains constrained by drug resistance and side effects, highlighting the demand for novel therapeutic agents. Cauloside A, a natural compound isolated from Fructus Akebiae, has shown anti-tumor potential; however, its precise mechanism of action in NSCLC has remained unclear. This study demonstrates that Cauloside A inhibits tumor growth by interacting with TLR4 to initiate a novel pyroptotic pathway. In A549 and H1299 cell lines, Cauloside A dose-dependently reduced cell viability and induced Pyroptosis, as evidenced by LDH release assays, transmission electron microscopy, and Annexin V/7-AAD staining. Mechanistic studies revealed that Cauloside A specifically activated GSDME cleavage via JNK phosphorylation and subsequent Caspase-3 activation, independent of GSDMD. Importantly, cellular thermal shift assays, surface plasmon resonance, and molecular docking analyses identified TLR4 as a potential binding target of Cauloside A. In a C57BL/6 mouse syngeneic (allograft) model, Cauloside A treatment resulted in dose-dependent suppression of tumor growth, accompanied by reduced Ki67 and PCNA expression without apparent toxicity. Western blot analysis confirmed consistent activation of the TLR4-JNK-caspase-3-GSDME signaling axis in tumor tissues. These findings establish Cauloside A as a potential TLR4 modular that triggers GSDME-dependent Pyroptosis, providing both mechanistic insights and a promising therapeutic strategy for NSCLC.