• Journal of Life Science
• /
• v.20 no.11
• /
• pp.1617-1624
• /
• 2010

Recently, it has been found that Asiasari radix showed a hypopigmenting effect on melanogenesis through activation of mitogen-activated protein kinase (MEK)/extracellular signal-activated kinase (ERK) in B16F10 melanoma cells. However, the hypopigmenting effect of A. radix on the $\alpha$-melanocyte stimulating hormone ($\alpha$-MSH)-stimulated melanogenesis has remained unknown. The purpose of this study was to investigate the inhibitory mechanism of the partially purified A. radix (PPAR)-induced hypopigmentating effects on $\alpha$-MSH-stimulated melanogenesis in B16F10 mouse melanoma cells. PPAR strongly inhibited tyrosinase activity and leads to decreased melanin synthesis in $\alpha$-MSH-stimulated B16F10 melanoma cells. PPAR also decreased the $\alpha$-MSH-induced over-expression of the melanogenic enzymes, tyrosinase, tyrosinase-related protein (TRP)-1, dopachrome tautomerase (Dct) and microphthalmia-associated transcription factor (MITF). We further showed that PPAR inhibits $\alpha$-MSH-induced melanogenesis via phosphorylation of MEK/ERK and PI3K/Akt, and that their activation was blocked by MEK inhibitors, PD98059 and PI3K inhibitors, LY294002 in $\alpha$-MSH-stimulated B16F10 melanoma cells. These results suggest that PPAR inhibits $\alpha$-MSH-induced melanogenesis by activation of MEK/ERK and PI3K/Akt through MITF degradation, which may lead to down-regulation of tyrosinase.

• 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.

• The Korea Journal of Herbology
• /
• v.28 no.3
• /
• pp.69-74
• /
• 2013

Objectives : Since hypopigmentation is known to increase the risk of skin cancer, melanogenesis in the skin needs to be regulated. Here, we evaluated the melanogenesis stimulatory effects of a modified Goagium herbal remedy (HR) and HR+ox bile (Bos taurus domesticus) extract (OBE) to address hypopigmentation disorders. Methods : B16F10 melanoma cells were treated with different dosages of HR and HR+OBE for 24 to 48 h after 1 h of 10 nM ${\alpha}$-melanocyte stimulating hormone (${\alpha}$-MSH). After the treatment, cell viability, tyrosinase activity, melanin synthesis and the expression of genes related to melanin synthesis were measured and the regulation of the ${\alpha}$-MSH signalling through cAMP responding element binding protein (CREB) was determined. Results : HR and HR+OBE with the ranges of $15{\sim}100{\mu}g/mL$ did not affect cell viability in melanoma cells. The 1 h treatment of HR+OBE (50 and $100{\mu}g/mL$) potentiated the phosphorylation of CREB by enhancing ${\alpha}$-MSH signaling and its 24 h treatment increased CREB expression. Consistent with CREB potentiation, their treatment for 24 h, the expression of microphthalmia-associated transcription factor (MIFT), tyrosinase, tyrosinase related protein (TRP)-1 and TRP-2 were increased in realtime PCR. Ultimately, the 48 h treatment of HR+OBE (50 and $100{\mu}g/mL$) increased tyrosniase activity and melanin contents in the melanoma cells in comparison to the control. Conclusions : HR+OBE (50 and $100{\mu}g/mL$) increases melanin synthesis in B16F10 melanoma cells via the stimulation of tyrosinase activity and expression of MIFT, tyrosinase, TRP-1 and TRP-2. HR+OBE can be used as the a possible treatment for hypopigmentation of the skin.

• Journal of the Korean Applied Science and Technology
• /
• v.34 no.2
• /
• pp.418-425
• /
• 2017

This study investigated antioxidation activity through the content of total polyphenol, that of flavonoid and DPPH radical scavenging activity and measured the cytotoxicity against B16F10 melanoma and inhibiting function of melanin biosynthesis to evaluate antioxidation activity and melanocyte effect of pueraria lobata root extract. As the results of study, it was recognized that the toxicity did not show against B16F10 melanoma and the increase of generating melanin was inhibited as the results of measuring the inhibition function of melanin biosynthesis after inducing the generation of melanin by ${\alpha}$-MSH against B16F10 melanoma cell. The high contents of polyphenol and flavonoid was found as the contents of pueraria lobata root extract increases and DPPH radical scavenging activity as the results of antioxidation activity. Through this study, it was recognized that pueraria lobata root extract has the feasibility that can be used as the material of cosmetics as it has the excellent effect of antioxidation activity and inhibiting the generation of melanin against melanocyte, low toxicity against skin cell and its safety against melanocyte of skin was found.

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

• The Journal of Korean Obstetrics and Gynecology
• /
• v.21 no.1
• /
• pp.83-98
• /
• 2008

Purpose: This study was performed to determine the inhibitory effect of Soyosangagamhwajae(SYG) on melanin synthesis in B16F10 mouse melanoma cell. Methods: The Inhibitory effects of Soyosangagamhwajae(SYG) on melanin synthesis were determined by in-vitro assay. To elucidate inhibitory effects of SYG on melanin synthesis, we determined the melanin release in B16F10 cell. And to investigate the action mechanism, we assessed the gene expression of tyrosinase, TRP-1, TRP-2. PKA, $PKC{\beta}$ in B16F10 cell. Results: 1. SYG significantly inhibited melanin-release in B16F10 cell. 2. SYG significantly inhibited mushroom tyrosinase activity in vitro. 3. SYG significantly suppressed the expression of tyrosinase in B16F10 cell. 4. SYG significantly suppressed the expression of TRP-1, TRP-2 in B16F10 cell. 5. SYG significantly suppressed the expression of PKA, $PKC{\beta}$ in B16F10 cell. Conclusion: From these results, it may be concluded that SYG has the antimelanogenetic effect.

• Journal of Life Science
• /
• v.21 no.2
• /
• pp.251-256
• /
• 2011

EGCG, catechins in green tea, is a kind of phytochemical. Through the regulation of signal pathways, EGCG has been known to show anti-oxidant and anti-tumor effects in cells. In this study, we investigated the apoptotic effects of EGCG through AMP-activated protein kinase (AMPK) signal pathways, including hypoxia inducible factor-1 alpha (HIF-$1{\alpha}$). The experiments were performed in B16F10 melanoma cells in a hypoxic state. AMPK is activated by ATP consumption such as nutrient deficiency, exercise, heat shock, etc. The activated AMPK that plays an important role as an energy sensor inhibits proliferation of cancer cells, as well as inducing apoptosis. HIF-$1{\alpha}$, the primary transcriptional regulator of the response to oxygen deprivation, plays a critical role in modulating tumor growth and angiogenesis in a hypoxic state. The apoptotic effects of EGCG were studied in B16F10 cells in a hypoxic state. The results show that EGCG inhibits the transcriptional activity of HIF-$1{\alpha}$ and induces apoptosis. These observations suggest that EGCG may exert inhibitory effects of angiogenesis and control tumor cell growth in hypoxic melanoma cells.

• Journal of Pharmacopuncture
• /
• v.10 no.1 s.22
• /
• pp.85-100
• /
• 2007

Objectives : This research was executed to verify antitumor effect and protective effect on doxorubicin(Doxo)-induced toxicity of Cultivated Wild Ginseng(CWG) and synergic effect of CWG with Doxo in B16/F10 melanomas-bearing C57BL/6 mice. Methods : To evaluate protective effect on doxorubicin(Doxo)-induced toxicity and enhancing effect on the antitumor activity of Doxo, CWG water extract(0.5 ml) was intraperitoneally injected for 10 days, in combination with intraperitoneal injection of Doxo(4 mg/kg) on days 12, 16, 19, to mice subcutaneously inoculated with $2{\times}10^6/ml$ B16/F10 melanoma cells. In order to investigate antitumor effect of CWG, CWG water extract(0.5 ml) was intraperitoneally injected for 10 days to mice subcutaneously inoculated with $2{\times}10^6/ml$ B16/F10 melanoma cells. Results : The body weights of melanoma-bearing mice increased following B16/F10 cells inoculation. In contrast, such an increase in body weights was significantly attenuated by Doxo administration. Whereas CWG inhibits the decrease in body weights induced by Doxo. The tumor volume and tumor weights of melanomas-bearing mice dramatically increased following B16/F10 cells inoculation, In contrast, such an increase in tumor volume and tumor weights were significantly attenuated by Doxo or CWG administration. But the synergic effect of CWG with Doxo was not observed. The reduction of cellularity of seminiferous epithelia, level of spermatogonium and spermatid induced by Doxo was recovered by CWG administration. BrdU labeling index of spermatogonium was remarkably decreased in Doxo group but was no change in CWG group. Whereas the incidence and intensity of BrdU labelled spermatocytes and spermatids were increased by CWG administration than those of Doxo group. Conclusions : The obtained results suggest that CWG have antitumor effect and protective effect on doxo-induced testicular toxicity. This effect might be mediated through the supplementation of vital energy.

• 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.

• Journal of Life Science
• /
• v.26 no.2
• /
• pp.259-264
• /
• 2016

Metastatic cancer is one of the main causes of cancer-related death since they rarely respond to available treatments. There is epidemiologic evidence that high garlic consumption decreases the incidence of cancer. Recent studies of our laboratory have revealed that a garlic-extracts is effective in suppressing metastasis. For experimental metastasis, C57BL/6 mice were injected intravenously with melanoma B16F10 cells in the tail vein, and were orally administered various concentrations (0, 50, 100 or 200 mg/kg body weight) of garlic hexane extract (GHE) for 21 days. The incidence and the area of the melanoma cell colony occupied by the poorly differentiated carcinoma were significantly lower in dose-dependent in 50, 100 and 200 mg/kg BW GHE - treated mice compared with control mice. In conclusion, the results of the present study show that GHE administration prevents lung metastasis in C57BL/6 mice.