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Melanoma is one of the most aggressive types of cancer. Showing a remarkable surge in the last 50 years, melanoma has been projected to be continuously rising in the future. Therapy for advanced type melanoma still becomes a challenge due to low response rate and survival in 10 years. Interestingly, connected with the survival, several epidemiological and preclinical studies had reported that vitamin D deficiency had an association with the progression in several cancer types. Feldman et al 2014 had reported, both in vivo and in vitro studies revealed antiproliferative, antiangiogenic, apoptosis, and differentiation induction effects of calcitriol in various types of cancers. Unfortunately, the study about the calcitriol (1,25(OH)2D3) effects toward melanoma is still limited and its mechanism remains unclear. In the present study, by utilizing B16-F10 melanoma cell line, we elaborate calcitriol antiproliferative effect using MTS Assay in a dose-dependent manner for 24 hours and potential calcitriol-induced apoptosis signaling pathway using Western Blot. We had observed that calcitriol may inhibit melanoma cell proliferation in IC50 of 93.8 ppm lead to the induction of apoptosis-related proteins such as caspase-3, caspase-9, poly (ADP-ribose) polymerases (PARP), mTOR, and HIF1α. These calcitriol-s effects reflect its potential capability as potent adjuvant therapy for melanoma. Taken together, calcitriol inhibited cell proliferation and induced cell death in B16-F10 cell. Thus, showed its potential to be adjuvant therapy for treating melanoma.