• Proceedings of the SCSK Conference
• /
• 2003.09b
• /
• pp.241-242
• /
• 2003

The effects of N-acetylphytospingosine(NAPS), one of the phytospingosine derivatives, on melanogenesis of B 16F 1 0 mouse melanoma cell lines were investigated. We assessed the effect of NAPS on the depigmentation of B16F10 cells. The melanin content of cells was significantly reduced by NAPS. We examined the inhibitory effect of NAPS on tyrosinase activity using L-dopa as a substrate and the results showed that tyrosinase activity was inhibited in a does-dependent manner. The mRNA level of tyrosinase as well as that of tyrosinase related protein-l (TRP-l) and tyrosinase related protein-2 (TRP-2) genes were not affected by NAPS based on a reverse transcription-polymerase chain reaction (RT-PCR) assay. We also performed a Western blotting analysis using anti-tyrosinase antibody. It showed that there is no change in tyrosinase protein level after treatment of NAPS. These results suggest that the depigmenting mechanism of NAPS in B16F10 melanoma cells involves inhibition of melanosomal tyrosinase activity, rather than the mRNA expression or protein level of tyrosinase.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.28 no.5
• /
• pp.506-511
• /
• 2014

Commelina communis Ledeb is a widely used medication for the treatment of antidiabetic, antioxidant and hypoglycemic agent in Korea. Alpha-melanocyte stimulating hormone (${\alpha}$-MSH) is a major factor to stimulate melanin synthesis in the skin. The purposes of this study was to investigate the inhibitory effects of extract from Commelina communis Ledeb (ECC) on ${\alpha}$-MSH-stimulated melanogenesis in B16F10 cells. ECC suppressed melanin synthesis and intracellular tyrosinase activity in B16F10 cells or ${\alpha}$-MSH-induced B16F10 cells in a dose dependent manner. In study on the melanogenic protein expressions, it had especially influence on expressions of tyrosinase and tyrosinase-related protein (TRP-1). Tyrosinase and TRP-1 expressions were gradually decreased in a dose-dependent. Additionally, the extract also decreased the ${\alpha}$-MSH-induced over-expression of tyrosinase and TRP-1. This results show that the anti-melanogenic activity of ECC is correlated with the suppression of tyrosinase and TRP-1 protein expressions in B16F10 cells.

• Journal of Life Science
• /
• v.29 no.9
• /
• pp.972-976
• /
• 2019

Polyopes affinis is a kind of red algae found in the South coast and near Jeju Island of Korea. The purpose of this study was to investigate the effects of Polyopes affinis ethanol extract (PAEE) on melanogenesis in ${\alpha}-MSH$ stimulated B16F10 melanoma cells. Melanoma cells were cultured for 72 hr treated with PAEE. Total melanin content and the activity of tyrosinase, a key enzyme in melanogenesis, were measured. When the melanin content in B16F10 melanoma cells was tested, PAEE was decreased in a dose-dependent manner: treatment with 100, 300, and $500{\mu}g/ml$ caused 25%, 30%, and 35% reduction, respectively. Treatment of 100, 300, and $500{\mu}g/ml$ of PAEE caused 6%, 12%, and 21% reduction of tyrosinase activities in B16F10 melanoma cells. Also, PAEE suppressed the expression of tyrosinase, tyrosinase-related protein-1, tyrosinase-related protein-2, and melanocyte-inducing transcription factor in B16F10 melanoma cells. A concentration of $500{\mu}g/ml$ of PAEE showed a greater decrease in tyrosinase activity, melanin content, and melanogenic enzyme protein expression. These results indicate that PAEE inhibits melanin synthesis and tyrosinase activity, and Polyopes affinis ethanol extract could be used as a functional whitening agent.

• Molecules and Cells
• /
• v.43 no.5
• /
• pp.479-490
• /
• 2020

Interleukin-9 (IL-9) is well known for its role in allergic inflammation. For cancer, both pro- and anti-tumor effects of IL-9 were controversially reported, but the impact of IL-9 on tumor metastasis has not yet been clarified. In this study, IL-9 was expressed as a secretory form (sIL-9) and a membrane-bound form (mbIL-9) on B16F10 melanoma cells. The mbIL-9 was engineered as a chimeric protein with the transmembrane and cytoplasmic region of TNF-α. The effect of either mbIL-9 or sIL-9 expressing cells were analyzed on the metastasis capability of the cancer cells. After three weeks of tumor implantation into C57BL/6 mice through the tail vein, the number of tumor modules in lungs injected with IL-9 expressing B16F10 was 5-fold less than that of control groups. The percentages of CD4⁺ T cells, CD8⁺ T cells, NK cells, and M1 macrophages considerably increased in the lungs of the mice injected with IL-9 expressing cells. Among them, the M1 macrophage subset was the most significantly enhanced. Furthermore, peritoneal macrophages, which were stimulated with either sIL-9 or mbIL-9 expressing transfectant, exerted higher anti-tumor cytotoxicity compared with that of the mock control. The IL-9-stimulated peritoneal macrophages were highly polarized to M1 phenotype. Stimulation of RAW264.7 macrophages with sIL-9 or mbIL-9 expressing cells also significantly increased the cytotoxicity of those macrophages against wild-type B16F10 cells. These results clearly demonstrate that IL-9 can induce an anti-metastasis effect by enhancing the polarization and proliferation of M1 macrophages.

• Journal of Ginseng Research
• /
• v.35 no.1
• /
• pp.86-91
• /
• 2011

Ginsenoside (G)-F1 is an enzymatic metabolite generated from G-Rg1. Although this metabolite has been reported to suppress platelet aggregation and to reduce gap junction-mediated intercellular communication, the modulatory activity of G-F1 on the functional role of skin-derived cells has not yet been elucidated. In this study, we evaluated the regulatory role of G-F1 on the cellular responses of B16 melanoma cells. G-F1 strongly suppressed the proliferation of B16 cells up to 60% at 200 ${\mu}g/mL$, while only diminishing the viability of HEK293 cells up to 30%. Furthermore, G-F1 remarkably induced morphological change and clustering of B16 melanoma cells. The melanin production of B16 cells was also significantly blocked by G-F1 up to 70%. Interestingly, intracellular signaling events involved in cell proliferation, migration, and morphological change were up-regulated at 1 h incubation but down-regulated at 12 h. Therefore, our results suggest that G-F1 can be applied as a novel anti-skin cancer drug with anti-proliferative and anti-migration features.

• Proceedings of the Plant Resources Society of Korea Conference
• /
• 2011.10a
• /
• pp.16-16
• /
• 2011

Melanogenesis is a well-known physiological response of human skin that may occur because of exposure to ultraviolet light, for genetic reasons, or due to other causes. In our effectors to find new skin lightening agents, acanthoic acid (AA) was investigated for its ability to inhibit melanogenesis. The effects of AA isolated from A.koreanumun the expression of $\alpha$-MSH-induced melanogenic factors (tyrosinase, tyrosinase related protein (TRP)-1, TRP-2 and MITF (microphthalmla-associated transcriptional factor)) were investigated in murine B16F10 melanoma cells. The results indicate that AA was an effective inhibitor of melanogenesis in B16F10 cells. To elucidate the mechanism of the effect of AA on melanogenesis, we performed Western blotting for melanogenic proteins. AA inhibited melanogenic factors (tyrosinase, TRP-1, TRP-2) expressions. In this study, we also confirmed that AA decreased the protein level of MITF proteins, which would lead to a decrease of tyrosinase and related genes in B16F10 melanoma cells. In order to apply AA to the human skin, the cytotoxic effects of the AA were determined by MTT assays using human keratinocyte HaCaT cells. Based on these results, we suggest that AA be considered possible anti-melanogenic agent and might be effective against hyperpigmentation disorders for the topical application.

• Journal of Life Science
• /
• v.19 no.1
• /
• pp.75-80
• /
• 2009

Melanogenesis is a physiological process that results in the synthesis of melanin pigments. Tyrosinase is a key enzyme for melanin biosynthesis, and hyperpigmentation disorders are associated with abnormal accumulation of melanin pigments, which can be improved by treatment with depigmenting agents. Among the possible melanin-reducing compounds, tyrosinase inhibitors are most promising for preventing and treating pigmentation disorder and are used as skin-whitening agents in the cosmetic industry. In the present study, the effects of fucoidan on melanogenesis and tyrosinase activity of B16F10 melanoma cells were investigated. Melanin synthesis and tyrosinase activity in B16F10 melanoma cells were decreased in a dose-dependent manner by fucoidan. Melanin production and tyrosinase activity in B16F10 melanoma cells stimulated by a-melanocyte stimulating hormone (a-MSH) were inhibited by fucoidan with a dose-dependent manner compared to control. Fucoidan inhibited tyrosinase activity of B16F10 melanoma cells with a dose-dependent manner as assessed by 3,4-dihydroxyphenylalanine (DOPA) staining. In conclusion, these findings indicate that fucoidan, which inhibit melanin synthesis and tyrosinase activity, is an effective skin-whitening agent.

• The Korea Journal of Herbology
• /
• v.34 no.2
• /
• pp.75-82
• /
• 2019

Objectives : In this study, various biological effects of Curcuma longa L. have been studied, however, beneficial effect of Curcuma longa L. in skin health remain still unclear. In this study, Curcuma longa L. water extract (CLE) was prepared. Inhibitory effect of CLE on melanin accumulation of B16F10 cells and expression levels of skin fibril-related proteins of human skin fibroblasts (HSF) were evaluated. Methods : The cytotoxic effect of CLE in B16F10 cells and HSF were examined by MTT assay. Inhibitory effect of CLE on the ${\alpha}-MSH-$ and IBMX-induced melanin accumulation and tyrosinase activity were evaluated in B16F10 cells. The expression levels of connective tissue growth factor (CCN2), Smad2, procollagen $1{\alpha}2$, collagen $1{\alpha}2$, and fibronectin in CLE-treated HSF were analyzed by western blotting. Results : The CLE treatment (concentrations 10 to $400{\mu}g/ml$) for 72 h did not affect to the B16F10 viability. However, 200 and $400{\mu}g/ml$ of CLE treatment for 24 h showed cytotoxic effect in HSF. Therefore, the concentrations 10, 50, and $100{\mu}g/ml$ of CLE were chosen in this study. The CLE treatment for 72 h dose dependently and significantly suppressed melanin accumulation and tyrosinase activity of B16F10 cells. In addition, the CLE treatment up-regulated expression levels of skin fibril-related proteins such as CCN2, Smad2, procollagen $1{\alpha}2$, collagen $1{\alpha}2$, and fibronectin. Conclusions : In conclusion, these results suggest that the CLE could be used as a natural material for skin health.

• Korean Journal of Plant Resources
• /
• v.32 no.4
• /
• pp.275-281
• /
• 2019

The pathological condition of excessive melanogenesis causing freckles, melasma, senile lentigo, pigmented acne scars, and cancer has a critical impact on the wellness of individuals. The mechanism of melanogenesis is related to the expression of melanogenic enzymes. Here, we evaluated the inhibitory effect of pine cone (Pinus densiflora) extracts on melanogenesis. P. densiflora, the Korean Red Pine, is the predominant tree species in the cool, temperate forests of northeast Asia, occurring in pure stands across Korea, Japan, and parts of northern China and Russia. P. densiflora leaves, pollen, and bark have been widely used for traditional medicine, or edible purposes. However, pine cones are rarely used as natural raw materials, although they contain many bioactive phytochemicals. The pine cone ethyl acetate fraction (PEF) showed no toxicity to B16F10 cells at a concentration of less than $100{\mu}g/mL$. PEF inhibited the expression of microphthalmiaassociated transcription factor (MITF), tyrosinase and tyrosinase-related factors in B16F10 cells treated with 3-Isobutyl1-methylxanthine (IBMX). These results suggest that pine cones can be used as an effective natural melanogenesis inhibitory agent.

• Biomedical Science Letters
• /
• v.26 no.3
• /
• pp.179-185
• /
• 2020

Adenine, a purine base, is a structural component of essential biomolecules such as nucleic acids and adenine nucleotides. Its physiological roles have been uncovered. Adenine suppresses IgE-mediated allergy and LPS-induced inflammation. Although adenine is known to inhibit lymphocyte proliferation, the effect of adenine to melamoma cells is not reported. Here, we investigated the growth inhibitory effects of adenine on B16-F10 mouse melanoma cells. Adenine suppressed the proliferation of B16-F10 cells in dose-dependent manner with the maximal inhibitory dose of 2 mM. Adenine treatment induced cell death molecular markers such as PARP and caspase 3 cleavages. Pan-caspase inhibitor z-VAD dramatically rescued the cell death molecular markers, cell proliferation recovered marginally. These results provide the possibility of adenine to be used as an anti-tumor agent.