Abstract: Objective To investigate the effects of clotrimazole on the growth of oral squamous cell carcinoma (OSCC). Methods OSCC-25 cells growing in log phase were treated with various doses of clotrimazole. The concentration of IC50, cell cycle and cell cycle related protein were examined. Results The concentration of clotrimazole for inhibiting OSCC was IC50, 8. 51 μmol/L. Clotrimazole induced cell cycle arrest in the G0-G1, cell cycle phase, with a concomitant decrease of cells in the G2-M and S-phase. Furthermore, clotrimazole significantly decreased the levels of cyclin D, cyclin E and CDK-4. Conclusions Clotrimazole inhibits the growth of OSCC.