• 대한화장품학회:학술대회논문집
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• 대한화장품학회 2003년도 IFSCC Conference Proceeding Book II
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• pp.241-242
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• 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.

• 동의생리병리학회지
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• 제28권5호
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• pp.506-511
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• 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.

• 생명과학회지
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• 제29권9호
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• pp.972-976
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• 2019

본 연구는 참까막살 에탄올 추출물이 천연 미백 소재로의 가능성을 알아보기 위해 멜라닌 함량, 세포내 tyrosinase 활성 측정 및 Western blotting 실험이 수행되었다. 참까막살 에탄올 추출물의 농도에 따른 MTT assay를 통해 세포 생존율 및 증식에 큰 영향을 미치지 않는 $500{\mu}g/ml$ 농도까지를 실험 조건으로 설정하였다. B16F10 melanoma cell의 melanin 생성에 미치는 영향을 측정한 결과 대조군에 비하여 ${\alpha}-MSH$만 처리한 경우 뚜렷하게 증가하였고, 참까막살 에탄올 추출물을 100, 300, $500{\mu}g/ml$의 농도로 각각 처리한 결과 ${\alpha}-MSH$만을 처리한 것에 비해 25%, 30%, 35%로 농도 의존적으로 감소하였다. Tyrosinase 활성도 100, 300, $500{\mu}g/ml$의 농도로 처리한 결과 ${\alpha}-MSH$만을 처리한 것에 비해 6%, 12%, 21%로 감소하였다. 참까막살 에탄올 추출물의 멜라닌 합성 관련 단백질 발현에 대한 영향을 확인하기 위하여 Western blot으로 미백관련 전사인자인 MITF, TRP-1, TRP-2, Tyrosinase 발현을 조사하였다. ${\alpha}-MSH$을 단독 처리한 경우에 각 단백질의 발현이 현저하게 증가하는 것을 확인 할 수 있었고 반면 참까막살 에탄올 추출물을 처리한 경우 농도 의존적으로 감소하는 것으로 나타났으며 특히 $500{\mu}g/ml$의 농도에서 매우 효과적으로 억제되었다. 본 연구 결과 참까막살 에탄올 추출물은 멜라닌 생합성에 관련된 유전자인 MITF, TRP-1, TRP-2, Tyrosinase의 발현을 억제하는 것으로 나타나, 향후 기능성 화장품 개발에 있어 효과적인 미백 기능성 해양 소재로 활용될 것으로 기대된다.

• Molecules and Cells
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• 제43권5호
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• pp.479-490
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• 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
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• 제35권1호
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• pp.86-91
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• 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.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2011년도 임시총회 및 추계학술발표회
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• pp.16-16
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• 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.

• 생명과학회지
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• 제19권1호
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• pp.75-80
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• 2009

미역, 다시마 등 갈조류의 추출물로서 생리활성성분으로 각광 받고 있는 fucoidan이 멜라닌 합성에 미치는 영양을 알아봄으로써 미백제로서의 개발 가능성 여부를 확인하지 위하여 B16F10 melnoma 세포를 이용하여 멜라닌 합성과 tyrosmase 활성도에 대한 실험의 결과 B16F10 melnoma 세포의 멜라닌 생성과 tyrosinase 활성도는 fucoidan의 농도가 증가함에 따라 농도 의존적으로 억제되었다. B16F10 melnoma 세포에 ${\alpha}$-MSH를 투여 후 멜라닌 생성과 tyrosinase의 활성도 역시 fucoidan의 농도가 증가할수록 멜라닌 생성과 tyrosmase 활성도가 억제되는 경향을 보였다. 또한 DOPA 염색의 결과 fucoidan은 농도 의존적으로 tyrosinase 활성도를 억제하였다. 이상의 결과 해조 추출 다당류인 furosinase은 B16F10 melanoma 세포의 멜라닌 합성과 tyrosinase 활성도 억제 작용을 나타내므로 미백 활성 물질로서의 가능성을 가지고 있는 것으로 생각된다.

• 대한본초학회지
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• 제34권2호
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• pp.75-82
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• 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.

• 한국자원식물학회지
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• 제32권4호
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• pp.275-281
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• 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.

• 대한의생명과학회지
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• 제26권3호
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• pp.179-185
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• 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.