• YAKHAK HOEJI
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• v.47 no.1
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• pp.31-36
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• 2003

To investigate the effect of protein kinase on melanin production via cAMP-dependent pathway, we measured the melanin amount and tyrosinase activity in B16 melanoma cells stimulated by alpha-melanocyte stimulating hormone (MSH), forskolin and 8-Br-cAMP. MSH, forskolin and 8-Br-cAMP significantly increased both melanin production and tyrosinase activity in B16 cells. Melanin production and tyrosinase activity by MSH are significantly inhibited by cyclic AMP-dependent protein kinase inhibitor (KT5720) and protein kinase C down-regulation treated with PMA. Bisindolmaleimide (1$\mu$M), protein kinase C inhibitor, significantly inhibited melanin production and tyrosinase activity stimulated by MSH, forskolin and 8-Br-cAMP with the following order of potency: MSH＞forskolin＞8-Br-cAMP. Tyrosine kinase inhibitor, genistein and DHC, significantly inhibited both, but the inhibitory effect was more potent in 8-Br-cAMP-stimulated B16 cells than MSH-stimulated cells. NFkB inhibitor (parthenolide) significantly inhibited melanin production and tyrosinase activity. Neither melanin production nor tyrosinase activity induced by MSH, forskolin and 8-Br-cAMP were affected by KN-62 (calmodulin-dependent protein kinase II inhibitor), PD098059 (mitogen-activated protein kinase inhibitor, MAPKK) and worthmannin (phosphatidylinositol 3-kinase inhibitor). These results suggest that both protein kinase C and tyrosine kinase are involved in melanin production by cyclic AMP-dependent pathway and NFkB pathway may play an important role in cyclic AMP-dependent melanin production in B16 melanoma cells.

• Advances in Traditional Medicine
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• v.1 no.2
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• pp.6-13
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• 2000

Melanin is specifically produced in melanocytes. The pathway for melanin biosynthesis is mainly controlled by tyrosinase. To estimate the inhibitory effect of melanin biosynthesis from 31 medicinal plants extracts, we tested the inhibitory effects of the tyrosinase promoter on B16 mouse melanoma cells. The result of this study demonstrated that Mori Radicis Cortex and Castena Fractus extracts only in tested medicinal plant extracts have high inhibitory effects on tyrosinase promoters with very low cytotoxicity on B16 mouse melanoma cells. Therefore, extracts of Mori Radicis Cortex and Castena Fractus were evaluated as very effective negative regulators of tyrosinase gene expression.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.22 no.2
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• pp.104-117
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• 2009

Objective : Melanin is one of the most important facor in skin color. Melanin protects human skin from ultraviolet radiation otherwise it causes melanin pigmentation. So this experiment is carried out for test whether Scutellaria baicalensis Georgi(SBG) inhibits melanin synthesis and tyrosinase activity in mouse B16 melanoma cells. Method : The melanin synthesis inhibition effects of SBG were examined by in vitro melanin production assay. We assessed inhibitory effects of SBG on melanin contents from B16F1 melanoma cell, on tyrosinase activity(cell and cell free system), effect of SBG on the expression tyrosinase, Microphthalmia-associated Transcription Factor(MITF), Extracellular signal-regulated Kinase(ERK). Result : SBG inhibited melanin synthesis induced $\alpha$-MSH($\alpha$-Melanin Stimulating Hormone) in B16F1. SBG inhibited tyrosinase activity and expression. And SBG down-regulates MITF and stimulated ERK activation in B16F1. Conclusion : According to above results, SBG was improved its suppression effect to the inhibition of melanin synthesis, tyrosinase activation, and tyrosinase promotor activation. So SBG is considered to be used for an strong source of skin whitening effect.

• YAKHAK HOEJI
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• v.59 no.4
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• pp.177-183
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• 2015

The mechanism of melanogenesis induced by cyclosporin A (CsA) was investigated in B16 melanoma cells. CsA stimulated the production of melanin in a dose-dependent manner in the cells. In addition, CsA increased intracellular $Ca^{2+}$ concentration in a dose-related fashion. Treatment with BAPTA/AM, an intracellular $Ca^{2+}$ chelator significantly inhibited the CsA-induced intracellular melanin synthesis. CsA profoundly induced $Cl^-$ efflux, which was significantly blocked by niflumic acid (NFA) and flufenamic acid (FFA), specific and nonspecific inhibitors of $Ca^{2+}$-activated $Cl^-$ channels (CaCCs), respectively. Furthermore, these inhibitors of CaCCs significantly inhibited the CsA-induced stimulation of melanin synthesis. Taken together, these results suggest that the activation of CaCCs may play an important role in the CsA-induced stimulation of melanin synthesis in B16 cells. These results further suggest that CaCCs may be a good target for the management of hyperpigmentation of the skin reported in the patients treated with CsA.

• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.5
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• pp.1304-1308
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• 2008

This study was performed to evaluate the effect of persimmon leaves extract on the reactive oxygen species (ROS) in cultured melanoma cells. The B16/F10 melanoma cells were treated with various concentrations of t-butyl hydroperoxide (t-BHP). And also, the effect of persimmon leaves (PL) extract on the cytotoxicity mediated by t-BHP was done on the cell viability, tyrosinase activity and melanogenesis by colorimetric assays. In this study, t-BHP decreased cell viability in dose-dependent manner and XTT90 and XTT50 values were measured at 10 and 35 uM of PL, respectively in these culture. And also, XTT50 value was assessed as a highly toxic effect on cultured melanoma cells by the toxic criteria. In the effect of PL extract on the t-BHP-mediated cytotoxicity, PL extract significantly increased the cell viability injured by t-BHP in cultured B16/F10 melanoma cells. PL also showed the decreased tyrosinase activity and melanogenesis. From these results, it is suggested that ROS such as t-BHP showed highly toxic effect on cultured melanoma cells, and also, PL extract inhibited the tyrosinase activity and melanogenesis in cultured melanoma cells injured by ROS.

• Journal of Acupuncture Research
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• v.18 no.1
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• pp.186-201
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• 2001

Purpose ; In order to study the effect of Houttuyniae herbal acupuncture on growth of melanoma B 16 and immunity in mice, the Control group with normal saline acupunchure after subcutaneous inoculation of B16BL6 cells, the Sample I with Houttuyniae herbal acupuncture manufactured by distilled water extraction method after subcutaneous inoculation of B16BL6 cells and the Sample II with Houttuyniae herbal acupuncture manufactured by alcohol extraction method after subcutaneous inoculation of B16BL6 cells were divided. Methods ; To evauluate the effect of Houttuynias herbal acupuncture on growth of melanoma B16 in mice, the weight of mouse melanoma, body weight, spleen index( $\sqrt(Weight of spleen/Body weight){\times}100$), lymphocytes numbers in mouse peripheral blood and spleen tissue, the percentage of CD4+ T cells, CD8+ and the ratio of CD4+/CD8+ in mouse peripheral blood and spleen tissue are measured. Results ; In study of the weight of mouse melanoma, Houttuyniae herbal acupuncture groups(Sample I, Sample II) showed statistically significant inhibitory effect. And in study of effect of reduction of change in body weight of mouse, Sample I showed statistically significant inhibitory effect, too. In study of reduction of spleen index, lymphocytes numbers in mouse spleen tissue, and the percentage of CD4+ and the ratio of CD4+/CD8+ T cells in mouse peripheral blood, Houttuyniae herbal acupuncture groups(Sample I, Sample II ) showed with statistically significant inhibitory effect. Also, Sample I showed inhibitory effect of reduction of the percentage of CD8+ T cells in mouse peripheral blood and spleen tissue and Sample II showed inhibitory effect of reduction of the percentage of CD4+ and the ratio of CD4+/CD8+ T cells in mouse spleen tissue with statistical significance. Conclusions ; The inhibitory effect of reduction of lymphocytes numbers in mouse peripheral blood and IL-2 productivity, Houttuyniae herbal acupuncture groups(Sample I, Sample II) didn't show significant effect.

• The Journal of Korean Medicine
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• v.26 no.1 s.61
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• pp.1-10
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• 2005

Objective: We examined the antimetastatic effect of Curcuma zedoaria Roscoe (CZ) on pulmonary metastasis of B 16 cells. Methods: For 6 weeks, Zedoariae Rhizoma made from dried CZ were dissolved in distilled water and administered to mice 2 weeks before they were injected with B]6 melanoma cells. Mice were given CZ at doses of 250 and 500 mg/kg, and were compared for lung weight, survival days, and NO production. Results: Intake of CZ throughout the experiment extended the average survival time. Intake after B16 cell injection slightly prolonged survival time, but intake before B]6 cell injection did not influence life span. We examined the effect of CZ on macrophage function by measuring NO production. After the macrophages were given CZ for 6 weeks, the amount of NO generated by the macrophages stimulated with LPS in culture medium increased. NO generated by the macrophages also served as a cytotoxic factor against B16 melanoma cells. B16 melanoma-conditioned medium reduced NO production by macrophages. However, CZ treatment reversed the reduction in NO production by the conditioned medium significantly. Conclusion : These findings may suggest that macrophage function-modulating activity by CZ appears to underlie its antimetastatic activity, which leads to a decrease in the number of lung metastatic surface nodules and the extension of life span.

• Biomedical Science Letters
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• v.26 no.3
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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.

• Korean Journal of Korean Medical Institute of Dermatology and Aesthetics
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• v.1 no.1
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• pp.5-15
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• 2005

The purpose of this study was to investigate the effect of the aqueous extract of the Astragalus membranaceous(AM). AM showed inhibitory effect on the tyrosinase activity using L-tyrosine as a substrate. Tyrosinase plays an important role in the process of melanin polymer biosynthesis. In vitro AM extract(1mg/ml) inhibited melanin biosynthesis and are useful for the material used in cosmetics. B16 mouse melanoma cells were cultured in different concentrations. The non-cytotoxicity of the plant extracts was confirmed by MTT assay.

• Natural Product Sciences
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• v.8 no.4
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• pp.183-185
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• 2002

To investigate skin whitening natural substances, the effects on melanogenesis by measuring the tyrosinase activity and the melanin contents of three hydrolysable tannins, $1,2,6-tri-O-galloyl-{\beta}-D-glucose$ (1), 2,3-(S)-HHDP-D-glucose (2) and pedunculagin (3) in B16 melanoma cells were examined. $1,2,6-Tri-O-galloyl-{\beta}-D-glucose$ (1), 2,3-(S)-HHDP-D-glucose (2) and pedunculagin (3) inhibited tyrosinase activity in B16 melanoma cells in a dose-dependent manner.