Inhibitory Mechanism of Bee Venom Against A431 Cutaneous Squamous Cell Carcinoma Cells Based on Label-Free Proteomics and Untargeted Metabolomics.

The incidence of cutaneous squamous cell carcinoma (cSCC) has been rising steadily in recent years, yet treatment options for advanced-stage patients remain limited. Bee venom (BV) possesses diverse biological activities including inhibiting tumor cell growth in various tumor models, but its underlying mechanism against cSCC A431 cells remains unclear. This study evaluated the antiproliferative effect of BV on A431 cells using the CCK-8 assay and explored its mechanism of action through label-free proteomics and untargeted metabolomics. BV inhibited A431 cell proliferation in a dose-dependent manner, with a half-maximal inhibitory concentration of 11.57 μg/mL. Proteomics analysis revealed that 118 differentially expressed proteins were mainly enriched in the p53 signaling pathway, mineral absorption, and base excision repair pathways. Metabolomics screening yielded 336 differentially expressed metabolites, primarily enriched in amino acid biosynthesis and metabolism. BV exerts its anti-A431 cell bioactivity potentially by synergistically regulating tumor-related signaling pathways, mineral homeostasis, and proteins involved in DNA repair processes, thereby disrupting amino acid biosynthesis and metabolism. But the transcriptomic data failed to confirm TP53 upregulation. These findings provide a theoretical basis for apipuncture and BV-based cSCC therapies.