Jiawei Guilu Erxian Decoction enhances CD8⁺ T-cell activation and cytotoxicity via JNK/c-JUN activation to suppress non-small cell lung cancer tumor growth in vivo

Abstract

Background The immunological effects of Jiawei Guilu Erxian Decoction (JGED) in non-small cell lung cancer (NSCLC) have not been fully defined. This study investigated whether JGED influences CD8⁺ T-cell activity and tumor growth. Methods The effect of JGED-containing serum on the viability of A549 and LLC cells was examined in vitro . An orthotopic LLC lung tumor model was established to assess tumor burden after JGED treatment. CD8⁺ T-cell infiltration and activation in tumor tissues were evaluated by immunofluorescence and flow cytometry. CD8⁺ T cells were exposed to JGED-derived serum and analyzed for proliferation and activation of CD8+ T cells. Proteomic profiling was performed to identify JGED-related protein changes in CD8⁺ T cells, and p-JNK and p-c-JUN levels were confirmed by Western blotting. Results JGED treatment was associated with smaller tumors and greater CD8⁺ T-cell infiltration but did not affect lung cancer cell proliferation in vitro . JGED-derived serum promoted the proliferation and activation of CD8+ T cells. Proteomic analysis revealed differences in protein profiles between treated and control CD8⁺ T cells, with enrichment of pathways linked to JNK signaling. The serum concentrations of phosphorylated JNK and c-JUN increased, whereas the inhibition of phosphorylated JNK and c-JUN reduced these changes. In coculture assays, tumor cell viability decreased and apoptosis increased when CD8⁺ T cells were incubated with JGED-derived serum, whereas inhibition of JNK weakened these effects. Conclusion JGED has a potential role in reducing tumor burden in vivo via increased CD8+ T-cell activity, although these findings require further clinical validation.