Effects Of Vitamin On Melanogenesis In A375Human Melanoma Cells

Objective: Vitamins such as vitamin C, vitamin E and vitamin B which haveantioxidation can affect melanin synthesis through mediate tyrosinase that became a newdirection for the treatment of skin pigments diseases. In this research, in order to explorethe possible mechanism for the treatment of pigment related diseases melasma and vitiligo,and the feasibility of the vitamins use in clinical treatment of melasma and vitiligo, weobserved the effects of melanin synthesis with vitamin C, vitamin E, vitamin B1, vitaminB6and vitamin B12in A375human melanoma cells.Methods: To determine the concentration of vitamins on A375human melanoma, wedetected the cell growth and proliferation of different concentrations vitamin C, vitamin E,vitamin B1, vitamin B6and vitamin B12by MTT. The cell cycle distribution was tested byflow cytometry. To observate the effects of vitamin on A375human melanoma cells, wedetected the tyrosinase activity by dopa oxidase assay, and measured the melanin contentby NaOH-assay.Results:1. MTT results revealed that: treated Vitamins,24h later, there are no statisticalsignificance (p>0.05) between the concentrations of VitC, VES, VitB1, VitB6and VitB12with the control group. After treated Vitamins for48h, compared with the control group,both VitC and VES could inhibit A375human melanoma cells growth (p<0.05),0.01μg/ml VitB1,0.01μ g/ml VitB6and0.01μ g/ml VitB12promoted the growth of A375human melanoma cells (p<0.05), and high concentration of VitB1(10μ g/ml,20μ g/ml),VitB6(10μ g/ml,20μ g/ml) and VitB12(10μ g/ml,20μ g/ml) inhibited the growth ofA375human melanoma cells (p<0.05), while the other groups compared with the control group had no statistical significance (p>0.05). Treated Vitamins for72h, VitC and VEScould significantly inhibit A375human melanoma cells growth (p<0.01);0.01μg/mlVitaminB1,0.01μg/ml VitaminB6and0.01μg/ml VitaminB12could significantly promotethe proliferation (p<0.05), but high-concentration VitaminB1, VitaminB6and VitaminB12could inhibit A375human melanoma cells growth (p<0.05).2. FCM analysis indicated that G2+S Phrase cell rate of0.05mg/ml Vitamin C and5.0μg/ml VES were lowly than that of control group(p<0.01);0.01μg/ml VitaminB1,0.01μg/ml VitaminB6and0.01μg/ml VitaminB12could significantly promote the G2+SPhrase cell rate (p<0.01).3. Dopa oxidase assay and NaOH-assay indicated that low-concentration VitaminB1(0.01μg/ml), VitaminB6(0.01μg/ml) and VitaminB12(0.01μg/ml) could improme thetyrosinase activity and melanin content than that of control group (p<0.05);0.05mg/mlVitamin C and0.05mg/ml Vitamin E could inhibit the melanin content and tyrosinaseactivity of A375human melanoma cells (p<0.05).Conclusion:1. Vitamin C and Vitamin E could inhibit the growth the activity of A375human melanomacells, and reduce the contents of melanin.2. VitaminB1, VitaminB6and VitaminB12could dual-directional regulte the growth,tyrosinase activity and the contents of melanin.