Noncalcemic 20-hydroxyvitamin D3 inhibits human melanoma growth in in vitro and in vivo models

Abstract

A novel pathway of vitamin D₃ (D₃) metabolism, initiated by C20-hydroxylation of D₃ by CYP11A1, has been confirmed to operate in vivo. Its major product, 20(OH)D₃, exhibits antiproliferative activity in vitro comparable to that of 1,25(OH)₂D_3, but is noncalcemic in mice and rats. To further characterize the antimelanoma activity of 20(OH)D₃, we tested its effect on colony formation of human melanoma cells in monolayer culture and anchorage-independent growth in soft agar. The migratory capabilities of the cells and cell-cell and cell-extracellular matrix interactions were also evaluated using transwell cell migration and spheroid toxicity assays. To assess the antimelanoma activity of 20(OH)D₃ in vivo, age-matched immunocompromised mice were subcutaneously implanted with luciferase-labelled SKMel-188 cells and were randomly assigned to be treated with either 20(OH)D₃ or vehicle (n=10 per group). Tumor size was measured with caliper and live bioimaging methods, and overall health condition expressed as a total body score scale. The following results were observed: (i) 20(OH)D₃ inhibited colony formation both in monolayer and soft agar conditions, (ii) 20(OH)D₃ inhibited melanoma cells in both transwell migration and spheroid toxicity assays, and (iii) 20(OH)D₃ inhibited melanoma tumor growth in immunocompromised mice without visible signs of toxicity. However, although the survival rate was 90% in both groups, the total body score was higher in the treatment group compared to control group (2.8 vs. 2.55). In conclusion, 20(OH)D₃, an endogenously produced secosteroid, is an excellent candidate for further preclinical testing as an antimelanoma agent.