• Biomolecules & Therapeutics
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• v.20 no.2
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• pp.165-170
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• 2012

Cell migration plays a role in many physiological and pathological processes. Reactive oxygen species (ROS) produced in mammalian cells influence intracellular signaling processes which in turn regulate various biological activities. Here, we investigated whether melanoma cell migration could be controlled by ROS production under normoxia condition. Cell migration was measured by wound healing assay after scratching confluent monolayer of B16F10 mouse melanoma cells. Cell migration was enhanced over 12 h after scratching cells. In addition, we found that ROS production was increased by scratching cells. ERK phosphorylation was also increased by scratching cells but it was decreased by the treatment with ROS scavengers, N-acetylcysteine (NAC). Tumor cell migration was inhibited by the treatment with PD98059, ERK inhibitor, NAC or DPI, well-known ROS scavengers. Tumor cell growth as judged by succinate dehydrogenase activity was inhibited by NAC treatment. When mice were intraperitoneally administered with NAC, the intracellular ROS production was reduced in peripheral blood mononuclear cells. In addition, B16F10 tumor growth was significantly inhibited by in vivo treatment with NAC. Collectively, these findings suggest that tumor cell migration and growth could be controlled by ROS production and its downstream signaling pathways, in vitro and in vivo.

• Proceedings of the PSK Conference
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• 2003.04a
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• pp.211.1-211.1
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• 2003

There are now increasing evidences that free radicals and reactive oxygen species are involved in a variety of pathological events. Flavonoids. a group of polypenolic compounds, are widespread in the human food supply. This study was carried out to investigate the antioxidant activity of these compounds. myriceitn and (+)-catechin on B 16Fl0. murine melanoma cell line in oxidative stress. Oxidative stress was induced by exposure to hydrogen peroxide. (omitted)

• Proceedings of the SCSK Conference
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• 2003.09b
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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.

• Korean Journal of Acupuncture
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• v.29 no.1
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• pp.117-130
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• 2012

Objectives : The purpose of this study was to investigate the melanogenesis inhibition effect of Saururus chinensis BAILL (SC) on in B16F10 melanoma cells. Methods : SC was fractionated ethanol extract by the hexane, ethyl acetate, butanol and water. We confirmed the inhibitory effect of tyrosinase activity and melanogenesis of all fraction samples. Results : Hexane fraction of Saururus chinensis BAILL (HSC), ethyl acetate of SC (ESC), and butanol of SC (BSC) were discovered to inhibit tysoinase activity and melanogenesis in the absence or presence of ${\alpha}$-MSH. However, water fraction of SC (WSC) did not affect tyrosinase activity and melanogenesis. In addition, all fractions did not inhibit the catalytic activity of cell-free tyrosinase from B16F10 melanoma cell lines. Conclusions : These results suggest that HSC, ESC and BSC reduce pigmentation by indirectly regulating tyrosinase.

• Journal of Life Science
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• v.34 no.2
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• pp.105-112
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• 2024

In this study, the various whitening activities of Inonotus obliquus were assessed for potential use as functional cosmetic materials. When measuring electron donating ability to confirm the antioxidant ability of I. obliquus extract, increased activity was observed as the concentration increased, and it was found an outstanding antioxidant capacity of 82.1% at a 1,000 ㎍/ml concentration. Also, the tyrosinase inhibitory effect, related to a whitening effect, was found to have inhibitory activity that increased in a concentration-dependent manner. The results of verifying the viability of melanoma cells (B16F10) using an MTT assay showed cell viability of more than 80% at concentrations below 100 ㎍/ml. Therefore, cell-related experiments were conducted at 25, 50, and 100 ㎍/ml concentrations. By measuring protein expression related to melanin synthesis via treating B16F10 cells with I. obliquus extract, it was confirmed that protein expression was inhibited in all factors, depending on the concentration. TRP-1 and MITF appeared by 40.1% and 64.2% amount of expression, respectively, at 100 ㎍/ml concentrations, and tyrosinase and TRP-2 were verified as having better protein expression inhibition than arbutin. In measuring mRNA expression related to melanin synthesis by treating B16F10 cells with I. obliquus extract, it was confirmed that mRNA expression was suppressed as the concentration increased. Accordingly, it was confirmed that I. obliquus extract has excellent whitening activity and could be used as a cosmetic material.

• Journal of Life Science
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• v.18 no.3
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• pp.298-303
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• 2008

Using a phage peptide library approach, we have isolated a peptide ligand that binds to transferrin receptor on the surface of human melanoma cell, B16F10. The library was first screened twice by recovering internalized phages and was further screened three times by competitively eluting transferrin receptor-specific phages with human transferrin among the phages bound to the cell surface. The peptides displayed by the selected phages were fused to translocation and catalytic domain of Pseudomonas exotoxin to prepare recombinant toxins. After estimating cytotoxicity of each recombinant toxin toward B16F10 cell, seven clones were selected. Sequence analysis revealed that one of the clones displayed a peptide which had a significant sequence homology with human transferrin. The peptide was chemically synthesized and was shown to be functional in delivering cytotoxic agents into B16F10 cell via interaction with transferrin receptor.

• Journal of Life Science
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• v.23 no.12
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• pp.1495-1500
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• 2013

The objective of this study was to evaluate the anti-melanogenic effects of Hizikia fusiforme (HF) fractions in ${\alpha}$-melanocyte stimulating hormone-induced B16F10 mouse melanoma cells. Ethanol extractions of Hizikia fusiforme (EEHF) were subjected to fraction by using dichloromethane (CFHF), ethyl acetate (EAFHF), butanol (BFHF), and water (WFHF). EEHF, CFHF, and EAFHF inhibited tyrosinase activity and melanin synthesis in B16F10 mouse melanoma cells in a dose-dependent manner. The melanin contents were inhibited by 40.5% and 33.2% in response to treatment with 50 ${\mu}g/ml$ of EEHF and CFHF, respectively. In addition, tyrosinase activities showed a 53.3% and 54.1% reduction in treatment with 50 ${\mu}g/ml$ of EEHF and CFHF. Western blotting analysis showed that EEHF, CFHF, and EAFHF inhibited tyrosinase, TRP-1, TRP-2, and MITF expression in a dose-dependent manner. In conclusion, these findings indicate that ethanol and dichloromethane fractions of Hizikia fusiforme, which inhibit melanin synthesis and tyrosinase activity, are effective skin-whitening agents.

• Herbal Formula Science
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• v.26 no.3
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• pp.223-236
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• 2018

Objectives : We compared the inhibitory effects of herb-combined remedies, which were recorded on "Yooam" prescription of Dongeuibogam, on cell migration and invasion, two critical cellular processes that are often deregulated during metastasis, in B16F10 melanoma cells. For this purpose, water extracts of Sipyukmiryukieum (SYMRKU), Danjacheongpitang (DJCPT), Cheongganhaeultang (CGHUT) and Jipaesan (JPS) were used. Methods : Cytotoxicity was assessed by an MTT assay. Wound healing and matrigel transwell assays were used to examine on B16F10 cell migration and invasion. The levels of mRNA and protein expression of matrix metalloproteinases (MMPs) and tissue inhibitor of metalloproteinases (TIMPs) were analyzed by RT-PCR and Western blotting. Results : Our data showed that DJCPT showed the strongest inhibitory effect among the four prescriptions in inhibiting cell motility of B16F10 melanoma cells within the concentration range that was not cytotoxic. The inhibitory potential of colony formation was higher in DJCPT and SYMRKU compared to the other two types of prescriptions, and the inhibitory effect of invasiveness is shown in order of DJCPT, SYMRKU, CGHUT and JPS. DJCPT, and SYMRKU strongly inhibited the activity and expression of MMP-2 and MMP-9, which are important mediators in cancer invasion, compared to CGHUT and JPS, and the increased expression of TIMP-1 and TIMP-2 was also more effective in these two prescriptions. In conclusion, DJCPT is expected to exhibit the most potent blocking effect on migration and invasion among four herb-combined remedies compared in B16F10 melanoma cells. Conclusion : Overall, the results of this study will be used as an important source to validate these prescriptions in animal models and to understand the mechanism of action of herbal remedies recorded in Dongeuibogam.

• Molecular & Cellular Toxicology
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• v.2 no.2
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• pp.81-88
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• 2006

Sphingolipid metabolites regulate many aspects of cell proliferation, differentiation, and apoptosis. In the present study, we have assessed the effects of the novel phytosphingosine derivative, N-acetylphytospingosine (NAPS), on the depigmentation of murine B16F10 melanoma cells, and have also attempted to identify the possible signaling pathway involved, in comparison with $C_{2}-ceramide$. NAPS and $C_{2}-ceramide$ both inhibited the growth of the B16F10 cells in a dose-dependent manner. Melanin content and tyrosinase activity were significantly reduced in response to treatment with NAPS and $C_{2}-ceramide$ at concentrations in a range between $1-5\;{\mu}M$. However, the levels of tyrosinase mRNA, as well as the levels of tyrosinase related protein-1 (TRP-1) and tyrosinase related protein-2 (TRP-2) genes and the level of tyrosinase protein remained unaffected by treatment with either NAPS or $C_{2}-ceramide$. We also attempted to determine the signaling pathway exploited by NAPS and $C_{2}-ceramide$. Interestingly, the phosphorylation of Akt/PKB at serine 473 by NAPS was reduced at the 5 minute mark, whereas $C_{2}-ceramide$ induced the phosphorylation of Akt/PKB at serine 473. Finally, Akt/PKB activity in the NAPS-treated cells was elevated in comparison with the untreated cells. LY294002, a specific PI3-K inhibitor which is located upstream of Akt/PKB, inhibited the phosphorylation of Akt/PKB, but induced an increase in melanin synthesis. These results suggest that the activation of Akt/PKB at serine 473 is related with the suppression of melanin production in the B16F10 mouse melanoma cells. Therefore, the mechanisms exploited by NAPS and $C_{2}-ceramide$ responsible for the depigmentation of B16F10 cells were concluded to involve the inhibition of melanosomal tyrosinase activity.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.31 no.1 s.49
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• pp.115-119
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• 2005

Methanol extract of plants in Jeju were investigated for biological properties related to whitening cosmeceuticals such as melanin contents on melanoma cell, mushroom tyrosinase activity inhibition. We found that extracts of leaves of Hypochoeris radicata, Solanum nigrum, Solidago serotica Gynostenmma pentaphyllum and Taxus cuspidata inhibit melanin synthesis in B16F10 melanoma cells. However they have no tyrosinase inhibitory activity.