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

• Published in: Food technology and biotechnology Vol. 62; no. 1; pp. 15 - 25
• Main Authors: Talapphet, Natchanok, Kim, Moon-Moo
• Format: Journal Article
• Language: English
• Published: Croatia Sveuciliste U Zagrebu 01.01.2024; Sveuciliste u Zagrebu, Prehramheno-Biotehnoloski Fakultet; University of Zagreb Faculty of Food Technology and Biotechnology
• Subjects: Aging; Assaying; B16F1 cells; Biosynthesis; Cell viability; Cells; Cytotoxicity; Enzymatic activity; Enzyme activity; Enzymes; Ethylenediaminetetraacetic acid; Fluorescent antibody technique; Free radicals (Chemistry); Gene expression; Genes; Hair; Hydrogen peroxide; Immunofluorescence; Mammalian cells; Melanin; melanogenesis; Melanoma; melanoma cells; Methionine; Microphthalmia-associated transcription factor; Organic chemicals; Organic chemistry; Original Scientific Papers; polyamine; Polyamines; Polymerase chain reaction; Proteins; Putrescine; Real time; Reductases; Senescence; Signal transduction; Skin; Transcription factors; Tyrosinase; South Korea; polyamine; melanogenesis; melanoma cells; putrescine; B16F1 cells
• ISSN: 1330-9862; 1334-2606
• DOI: 10.17113/ftb.62.01.24.8120

Ageing is a biochemical, metabolic and genetic physiological phenomenon. The suppression of melanin biosynthesis, evident in the greying of the hair, is a hallmark of ageing 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 and plays a crucial role in regulating skin melanogenesis. Therefore, the purpose of this study is to explore the effect of putrescine on the signalling pathways of melanogenesis in melanoma cells. Melanin production capacity of putrescine was analysed using a tyrosinase activity assay. To assess the cell viability of B16F1 cells exposed to putrescine, a tetrazolium dye MTT assay was performed. The effect of putrescine on melanin synthesis in the presence of H O was evaluated using various assays in B16F1 cells. The effect of putrescine on melanin production in B16F1 cells was determined using a specific melanin production assay. Gene expression was analysed using real-time polymerase chain reaction (RT-PCR). Furthermore, the effect of putrescine on the expression of proteins related to melanin production in the cells treated with H O was analysed by immunofluorescence and Western blot analysis. Putrescine increased tyrosinase activity and showed no cytotoxicity in B16F1 cells. In addition, putrescine effectively scavenged H O , as shown by the reduction of intracellular H O amounts in 2',7'-dichlorofluorescin diacetate analysis, and promoted melanin production in living cells. The stimulation of melanogenesis by putrescine was attributed to the increased expression of , , and genes. Immunofluorescence assays revealed that putrescine enhanced the expression of proteins associated with melanogenesis and upregulated TYR, TRP-1 and TRP-2 the microphthalmia-associated transcription factor (MITF) and increased the expression of methionine sulfoxide reductases A (MSRA) and B (MSRB) in the cells treated with H O , effectively promoting melanogenesis. These results suggest that putrescine can be used to stimulate melanin synthesis. This is the first study to investigate the effect of putrescine on the signalling pathways of melanogenesis in B16F1 melanoma cells. The results confirm that putrescine can promote melanogenesis through the expression of TYR, TRP-1 and TRP-2 the MITF in cells treated with H O . Putrescine can be used exclusively as a cosmetic product to prevent premature greying of hair.