Putrescine Upregulates Melanogenesis Through Modulation of MITF Transcription Factor in B16F1 Mouse Melanoma Cells

Abstract

Research background. Aging is a biochemical, metabolic, and genetic physiological phenomenon. The suppression of melanin biosynthesis, evident in the graying of hair, is a hallmark of aging resulting from translation failure, reduced enzyme activity, and cellular senescence. Putrescine, the smallest member of the polyamine family and an organic chemical, is present in living mammalian cells, playing a crucial role in regulating skin melanogenesis. Therefore, the purpose of this study is to explore the effect of putrescine on the signaling pathways of melanogenesis in melanoma cells. Experimental approach. Putrescine was studied on the melanin production capacity was examined through a tyrosinase activity assay. To assess the cell viability of B16F1 cells exposed to putrescine, an MTT assay was performed. The impact of putrescine on melanin synthesis in the presence of H2O2 was evaluated using various in vitro assays in B16F1 cells. The effect of putrescine on melanin production in B16F1 cells was achieved through a dedicated melanin production assay. Gene expression analysis was conducted using RT-PCR. Furthermore, the impact of putrescine on the expression of proteins related to melanin production in H2O2-treated cells was examined through immunofluorescence and western blot analysis. Results and conclusions. Putrescine increased tyrosinase activity and demonstrated non-cytotoxicity in B16F1 cells. Furthermore, putrescine effectively scavenged H2O2, as evidenced by the reduction in intracellular H2O2 levels in DCFH-DA analysis, and promoted melanin production in living cells. The stimulation of melanogenesis by putrescine was attributed to the elevated expression of Mitf, Tyr, Trp-1, and Trp-2 genes. Immunofluorescence investigations revealed that putrescine enhanced the expression of proteins associated with melanogenesis and upregulated TYR, TRP-1, and TRP-2 via the MITF transcription factor and increased the expression of MSRA and MSRB in H2O2-treated cells, thereby effectively promoting melanogenesis. These findings suggest that putrescine may be utilized to stimulate melanin synthesis. Novelty and scientific contribution. Putrescine could be solely used as a cosmetic agent to prevent premature graying of hair.