• Journal of Life Science
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• v.24 no.9
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• pp.1001-1005
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• 2014

Ginsenoside Rg3 is one of the active ingredients extracted from red ginseng, and it is an effective chemical component of the human body and well known in herbal medicine as a restorative agent. Several studies have shown that Rg3 has a potent anti-tumor effect on various cancer cell lines. However, Rg3-induced apoptosis in B16F10 melanoma cancer cells is not well understood. In the present study, we tested whether ginsenoside Rg3 could induce apoptosis in B16F10 melanoma cells. We found that Rg3 could inhibit B16F10 melanoma cell viability in a dose-dependent manner, but not normal cells, such as EA.hy.926 and NIH3T3 cells. We also found that Rg3 could induce apoptosis in B16F10 melanoma cells using tunnel-staining assay in a dose-dependent manner. Rg3 treatment induces the phosphorylation of p38 and the expression of Bax, but it inhibits the expressions of the phosphorylation of focal adhesion kinase Bcl2 and pro-caspase3. Taken together, our data suggest that Rg3 could be useful as an anti-cancer agent in B16F10 melanoma cells.

• Korean Journal of Food Science and Technology
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• v.54 no.2
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• pp.134-140
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• 2022

Kelp belongs to the brown algae family and has been reported to exert anti-cancer effects on some cancer types, however studies have not been reported on the anti-cancer effects of kelp extracts on melanoma. In this study, the anti-cancer effects of kelp extract in B16-F0 cells were investigated, and the underlying molecular mechanisms were assessed. Kelp extract was found to inhibit the proliferation of B16-F0 cells, induce cytotoxicity, inhibit cell colony formation, and induce DNA fragmentation and apoptosis. The molecular mechanism was found to involve kelp extract increasing the expression of cytochrome-c and activated caspase-9 in the intrinsic apoptotic pathway. In addition, kelp extract upregulated the expression of Fas-associated protein with death domain and activated caspase-8 in the extrinsic apoptosis pathway. Activation of caspase-9 and caspase-8 by kelp extract induced activation of caspase-3 and cleaved poly adenosine diphosphate-ribose polymerase, consequently inducing apoptosis. These data suggest that kelp extract represents a potential therapeutic agent for melanoma.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2018.04a
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• pp.63-63
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• 2018

본 연구에서는 80% 식물성 알코올을 추출 용매로 사용해 오디를 빛을 차단 후 실온에서 3일 간 추출하였다. 3회 여과한 후 최소 온도($40{\sim}60^{\circ}C$)에서 농축한 뒤 동결 건조하여 파우더 형태로 사용하였다. 오디(Morus alba)의 에탄올 추출물은 B16/F10 세포의 항산화 및 멜라닌 합성 억제 효과를 나타내었다. 멜라닌 함량과 세포 내 tyrosinase 활성을 Western blotting으로 측정 하였다. Tyrosinase와 tyrosinase-related protein (TRP) -1은 tyrosinase-related protein (TRP) -2보다 강력하게 억제되었으며, 이들 결과는 tyrosinase와 TRP-1은 흑갈색을 띠는 eumelanin의 생합성의 억제와 강한 상관관계가 있음을 보여 주었다. ${\alpha}$-melanocyte-stimulating hormone (${\alpha}$-MSH) 처리 한 B16/F10 흑색 종 세포에서 M. alba 에탄올 추출물은 멜라닌 생성 연관 단백질의 발현 및 멜라닌 생성이 용량 의존적으로 억제 하였다. 멜라닌 함량과 세포 내 tyrosinase 활성을 Western blotting으로 측정 하였다. 또한 DPPH와 SOD를 사용하여 항산화 활성을 분석하였고 총 폴리 페놀과 총 플라보노이드 함량을 측정 하였다. MTT assay 분석을 사용하여 M. alba 에탄올 추출물의 세포 독성을 측정 하였다. B16/F10 멜라닌 생성 세포의 tyrosinase 저해 활성 및 사멸 효과가 일반적으로 효과적이었다. 따라서 M. alba 에탄올 추출물은 항산화 및 미백 효과를 나타내며, 기능성 화장품의 천연 성분으로서 우수한 것으로 여겨진다.

• The Journal of Internal Korean Medicine
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• v.33 no.4
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• pp.405-417
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• 2012

Objectives : In the present study, anti-metastatic properties of the methanol extract of L. obtusiloba (MLO) were evaluated. Methods : To determine the effect of MLO on cancer metastasis, we checked matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) activities and expressions in B16F10 melanoma cells. In addition, we performed cell migration assay as well as invasion assay using Matrigel. Finally, we used an in vivo lung metastasis model to confirm the anti-metastatic activity of MLO. Results : 1. MLO showed potent inhibitory effects on MMP-2 and MMP-9 activities and expressions via down-regulation of activation of NF-${\kappa}B$ in B16F10 melanoma cells. 2. Melanoma cell migration and invasion were down-regulated by MLO treatment. 3. Not only in vitro model, but MLO also significantly suppressed lung metastasis in vivo. Conclusions : The present results indicate that MLO has strong inhibitory effect on cancer metastasis. Therefore, L. obtusiloba could be a valuable anti-metastatic agent.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.47 no.2
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• pp.107-121
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• 2021

Skin hypopigmentation, which is observed in albinism or vitiligo, occurs when melanin synthesis is decreased by genetic, epigenetic, and other factors. To identify drug candidates that can promote melanin synthesis in cells, we screened an epigenetic modulator library consisting of 141 cell-permeable, small molecule drugs. B16/F10 murine melanoma cells were treated with each drug at 0.1 𝜇M and melanin synthesis and cell viability were subsequently monitored. As a result, (-)-neplanocin A, 3-deazaneplanocin A (DZNep), and DZNep hydrochloride were found to increase cellular melanin synthesis without causing cytotoxicity. Because these three structurally related drugs exhibited similar dose-dependent effects on melanin synthesis and cell viability, DZNep was selected as a representative drug for additional experiments. DZNep increased intracellular melanin content and tyrosinase (TYR) activity. DZNep also induced the expression of TYR, tyrosinase-related protein 1 (TYRP1), and dopachrome tautomerase (DCT) at the mRNA and protein levels. DZNep also induced the mRNA and protein expression of microphthalmia-associated transcription factor (MITF), a key regulator of melanin synthesis. DZNep is a specific inhibitor of S-adenosylhomocysteine hydrolase and it caused the accumulation of S-adenosylhomocysteine that inhibits histone methyltransferases in cells. This study suggests that melanogenesis can be modulated by targeting S-adenosylhomocysteine hydrolase in certain cellular contexts.

• Journal of Life Science
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• v.32 no.5
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• pp.331-339
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• 2022

Melanin is a pigment produced by melanocytes to protect the skin from external stimuli, mainly ultraviolet (UV) rays. However, abnormal and excessive production of melanin causes hyperpigmentation disorders, such as freckles, age spots, and discoloration. Natural cosmeceuticals are a new trend for treating or preventing hyperpigmentation due to fewer side effects and biocompatibility. In this context, the current study focused on Cnidium japonicum, a halophyte with several uses in folk medicine, to evaluate its potential as a skin-whitening agent. The effect of C. japonicum extract (CJE) on melanin production was analyzed in melanogenesis-stimulated B16F10 melanoma cells. The results showed that CJE successfully inhibited the oxidation of tyrosine and L-DOPA by tyrosinase and subsequently decreased the production of the key enzymes responsible for melanin production: tyrosinase, tyrosinase-related protein-1, and protein-2. This effect was confirmed by decreased intracellular and extracellular melanin levels in B16F10 melanoma cells after CJE treatment. Further experiments to elucidate the action mechanism revealed that CJE treatment suppressed melanin production by inhibiting the activation of glycogen synthase kinase 3 β (GSKβ)/β-catenin and protein kinase A (PKA)/cAMP-response element binding protein (CREB) pathways, which are the upstream activators of melanogenesis. In conclusion, the present study suggests that C. japonicum is a potential natural source of bioactive substances for the development of novel cosmeceuticals that can act against hyperpigmentation.

• Journal of Life Science
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• v.17 no.6 s.86
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• pp.873-875
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• 2007

Effects of hot-water extract from Sasa quelpaertensis leaf (HWES) on melanogenesis were investigated in B16/F10 mouse melanoma cells. HWES inhibited cellular tyrosinase activity and melanin biosynthesis in a dose-dependent manner. Western blotting analysis showed that HWES dose-dependently inhibited tyrosinase and tyrosinase related protein-1 expression. Also, HWES suppressed sustained ERK activation in a concentration-dependent manner, suggesting that HWES inhibits the melanin biosynthesis through the suppressive effect against pathway involving sustained ERK activation.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.5
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• pp.705-711
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• 2014

In order to develop a skin-whitening agent, melanin contents and intracellular tyrosinase activity were determined by western blotting. Ethyl acetate fractions of 80% ethanol extracts from lily (Lilium Oriental Hybrid 'Siberia') bulbs (R-EA) inhibited melanin synthesis in a dose-dependent manner in ${\alpha}$-melanocyte stimulating hormone (${\alpha}$-MSH)-treated B16/F10 murine melanoma cells. Intracellular tyrosinase activity and melanin contents were suppressed by 45% and 74%, respectively, in response to treatment with $100{\mu}g/mL$ of R-EA. Examination of protein expression associated with ${\alpha}$-MSH-induced melanogenesis showed that tyrosinase related protein (TRP)-1 was inhibited more strongly than tyrosinase, and these results were correlated with stronger inhibition of melanin synthesis than intracellular tyrosinase activity. Taken together, R-EA containing p-coumaric acid and resveratrol could be used as a hypopigmentation agent through suppression of sustained extracellular signal-regulated kinase (ERK) activation via melanogenic induction.

• 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.29 no.11
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• pp.1200-1207
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• 2019

Ultraviolet radiation exposure is a major cause of extrinsic skin aging, which leads to skin hyperpigmentation. Loganin, a major iridoid glycoside obtained from Corni fructus, has anti-inflammatory, anti-diabetic, and neuroprotective effects. In this study, we investigated the mechanisms underlying the anti-melanogenic effects of loganin in B16F10 melanocytes treated with ${\alpha}$-melanocyte stimulating hormone (${\alpha}-MSH$) and 3-isobutyl-1-methylxanthine (IBMX). Anti-melanogenic activity was measured by treating cells with loganin at concentrations between 1 and $20{\mu}m$. Cell viability assays confirmed that doses of loganin up to $20{\mu}m$ were not cytotoxic. Loganin significantly and dose-dependently decreased intracellular melanin production. We also investigated potential molecular signaling pathways for the anti-melanogenesis effects of loganin. Western blotting showed that treatment with ${\alpha}-MSH$ and IBMX increased the phosphorylation of cAMP response element-binding protein (CREB) and the gene expressions of microphthalmia-associated transcription factor (MITF) and tyrosinase. Addition of loganin suppressed these increases, while promoting the phosphorylation of extracellular signal regulated kinase (ERK) and the anti-melanogenesis response. Our data therefore indicated that loganin could attenuate the increased melanin synthesis induced by ${\alpha}-MSH$ and IBMX treatment of B16F10 melanocytes. This attenuation appears to occur by downregulation of CREB phosphorylation and MITF and tyrosinase gene expression and upregulation of ERK phosphorylation. These finding suggests that loganin could be a valuable candidate for treatment of skin diseases related to hyperpigmentation.