Oral squamous cell carcinoma (OSCC) is a highly aggressive malignancy with poor prognosis due to late detection, rapid progression, and frequent metastasis, underscoring the urgent need for novel therapeutic targets. This study investigates the roles of kinesin family member 2C (KIF2C) and Polo-like kinase 1 (PLK1) in OSCC progression and their functional interplay. Immunohistochemical and western blot analyses revealed marked upregulation of KIF2C and PLK1 in human OSCC tissues and cell lines (SCC9, SCC25, Cal27). Functional characterization in Cal27 cells (selected for highest KIF2C expression via qPCR/WB) demonstrated that KIF2C knockdown via siRNA transfection suppressed proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT), while inducing apoptosis and G0/G1 cell cycle arrest. Mechanistically, KIF2C silencing downregulated PLK1 expression, concomitantly reducing EMT markers (N-cadherin, vimentin), matrix metalloproteinases (MMP-2/9), and angiogenesis factors (VEGF, α-SMA). Complementary assays (CCK-8, EdU, Transwell, wound healing) and flow cytometry confirmed that KIF2C-PLK1 axis promotes tumor growth by enhancing matrix degradation, angiogenesis, and S-phase proliferation while inhibiting apoptosis. These findings establish KIF2C as a pivotal regulator of OSCC progression through PLK1-mediated signaling, highlighting their dual potential as prognostic biomarkers and therapeutic targets for OSCC management.