Background: Oral cancer ranks as the sixth most common malignancy worldwide, with a significantly higher prevalence among men aged 40 to 60 years than women. Previous studies have demonstrated that periplocin exhibits therapeutic potential by inhibiting cancer cell proliferation and inducing apoptosis across various cancer types. However, its role in oral squamous cell carcinoma (OSCC) and the associated molecular mechanisms remain insufficiently understood. In this study, we aimed to investigate the inhibitory effects of periplocin on the malignant behaviors of OSCC cells. Methods: MTS [3-(4,5-dimethyl-2-thiazolyl)-2 ,5-diphenyl-2-H-tetrazolium bromide] assay, colony formation assay, flow cytometry, and transwell assay were employed to assess the effects of periplocin on OSCC cell proliferation, apoptosis, cell cycle progression, and migration. Furthermore, proteomics analysis was conducted to identify differentially expressed proteins in OSCC cells treated with periplocin, providing insights into its mechanisms of action. Results: Periplocin significantly influenced OSCC cell proliferation, apoptosis, cell cycle dynamics, and migration. Proteomics data indicated that periplocin modulates apoptosis, protein binding, and DNA replication signaling pathways. Furthermore, bioinformatics analysis revealed a strong association between syndecan 1 (SDC1) and coiled-coil-helix-coiled-coil-helix domain containing 2 (CHCHD2) in OSCC. Conclusions: The findings of this study indicate that periplocin exerts anti-OSCC effects by modulating key cellular processes in OSCC cells, offering a promising therapeutic avenue for OSCC management.