OBJECTIVE The inverse association between vitamin D and cancer risk is well-established, but the relationship with oral cancer is less well-understood. To further the understanding of these relationships, this study sought to evaluate any growth-inhibiting effects of vitamin D on well-characterized oral cancers. METHODS This study utilized 1,25-dihydroxy Vitamin D3 to evaluate any changes in growth using CAL27, SCC15, and SCC25 oral cancer cell lines at physiological and supraphysiological concentrations. RESULTS These assays revealed that the growth of all three cancer cell lines was significantly reduced by vitamin D administration, with maximal inhibition in SCC15 of -6.8% at 50 nmol, -19.7% in CAL27, and -43.6% in SCC25 at 100 nmol (p < .05). In addition, the observed decreases in growth were associated with significant decreases in viability (ranging from -18% in SCC15, -23% in CAL27, and -47% in SCC25 cells), as well as activation of two key apoptotic pathways (caspase and bcl:bax). CONCLUSIONS The results of this study demonstrate the growth-inhibitory effects of vitamin D administration in specific oral cancer cell lines, which will enhance the understanding of oral oncologists and oral health researchers in developing standards for generalizing the health-protective effects of diet and dietary supplements as treatment options for patients with oral cancer.