• Journal of Applied Biological Chemistry
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• v.63 no.1
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• pp.89-93
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• 2020

In this study, the efficacy of melanoma cell B16F10 was investigated using the Korean native plant Oplismenus undulatifolius (OU). First, the cell viability of the extract was more than 90% when treated with 15 ㎍/mL of phenolics from OU. The results showed that melanin biosynthesis and cellular tyrosinase synthesis were inhibited by treatment with α-melanocyte-stimulating hormone-stimulated mouse melanoma cell B16F10 at a concentration of 15 ㎍/mL of phenolics for cell-line efficacy. The expression of tyrosinase, tyrosinase-related protein (TRP)-1, TRP-2, and microphthalmia transcription factor (MITF) protein was confirmed by western blot to investigate the effect of phenolics from OU on melanin biosynthesis. When treated with phenolics from OU 15 ㎍/mL, tyrosinase, TRP-1, TRP-2, and MITF decreased the protein expression level. In particular, tyrosinase, TRP-1, and MITF inhibited the production amount to a level similar to that of the non-treated normal group, indicating that the effect was excellent. Therefore, phenolics from OU acts as an inhibitor of tyrosinase, TRP-1, TRP-2, and its transcription factor MITF, and participates in melanin biosynthesis mechanism. These results suggested the potential for development as a material.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.7
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• pp.3519-3528
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• 2012

Inhibitory effects of Maclura amboinenesis Bl, one plant used traditionally for the treatment of cancers, on metastatic potential of highly metastatic B16F10 melanoma cells were investigated in vitro. Cell proliferation was assessed using the MTT colorimetric assay. Details of metastatic capabilities including invasion, migration and adhesion of B16F10 melanoma cells were examined by Boyden Chamber invasion and migration, scratch motility and cell attachment assays, respectively. The results demonstrated that n-hexane and chloroform extracts exhibited potent anti-proliferative effects (p<0.01), whereas the methanol and aqueous extracts had less pronounced effects after 24 h exposure. Bioactivity-guided chromatographic fractionation of both active n-hexane and chloroform extracts led to the isolation of two main prenylated xanthones and characterization as macluraxanthone and gerontoxanthone-I, respectively, their structures being identified by comparison with the spectral data. Interestingly, both exhibited potent effective effects. At non-toxic effective doses, n-hexane and chloroform extracts (10 and $30{\mu}g/ml$) as well as macluraxanthone and gerontoxanthone-I (3 and $10{\mu}M$) significantly inhibited B16F10 cell invasion, to a greater extent than $10{\mu}m$ doxorubicin, while reducing migration of cancer cells without cellular cytotoxicity. Moreover, exposure of B16F10 melanoma cells to high concentrations of chloroform ($30{\mu}g/ml$) and geratoxanthone-I ($20{\mu}M$) for 24 h resulted in delayed adhesion and retarded colonization. As insights into mechanisms of action, typical morphological changes of apoptotic cells e.g. membrane blebbing, chromatin condensation, nuclear fragmentation, apoptotic bodies and loss of adhesion as well as cell cycle arrest in the G1 phase with increase of sub-G1 cell proportions, detected by Hoechst 33342 staining and flow cytometry were observed, suggesting DNA damage and subsequent apoptotic cell death. Taken together, our findings indicate for the first time that active n-hexane and chloroform extracts as well as macluraxanthone and gerontoxanthone-I isolated from Maclura amboinensis Bl. roots affect multistep of cancer metastasis processes including proliferation, adhesion, invasion and migration, possibly through induction of apoptosis of highly metastatic B16F10 melanoma cells. Based on these data, M. amboinensis Bl. represents a potential candidate novel chemopreventive and/or chemotherapeutic agent. Additionally, they also support its ethno-medicinal usage for cancer prevention and/or chemotherapy.

• Journal of Applied Biological Chemistry
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• v.61 no.3
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• pp.267-273
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• 2018

Matricaria chamomilla L. has been used as a bath agent in Europe because of its sterilization effect on the skin. Flowers contain terpenes, flavonoids are effective in relieving inflammation. Matricaria chamomilla L. has been reported to have various drug efficacies such as sedation, anti-diabetic effect and anti-arthritic effect, but there is little research on the scientific efficacy of whitening effect. The purpose of this study was to examine the whitening effect of Matricaria chamomilla L. extract and to investigate the mechanism of inhibition of melanogenesis. The extracts were used to determine tyrosinase inhibitory activity. The tyrosinase inhibitory activity of extracts was ineffective for water extract but in 60% ethanol extract was shown in a concentration-dependent manner. B16F10 melanoma cell was measured using a powder obtained by lyophilization of 60% ethanol extract. The toxicity was observed at a concentration of $75{\mu}g/mL$. And concentration range was selected to be at most $50{\mu}g/mL$. The effect of tyrosinase, MITF, TRP-1 and TRP-2 on the expression of melanin protein was investigated in melanoma cells of B16F10 melanoma cells. As a result, it was confirmed that as the concentration of the extract increased, the melanogenesis level decreased and the protein expression level also decreased in a concentration dependent manner. Therefore, it was concluded that Matricaria chamomilla L. extract inhibited melanogenesis in cells. Based on the above results, it is expected that it will be used as a useful basic data for industrialization of whitening functional food of Matricaria chamomilla L.

• IMMUNE NETWORK
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• v.5 no.3
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• pp.163-171
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• 2005

Background: To perform the successful dendritic cell-based cancer immunotherapy one of the main issues to be solved is the source of antigen for DC pulsing. Limitations occur by using auto-tumor lysate due to the difficulties obtaining enough tumor tissue(s) quantitatively as well as qualitatively. In this study the possibility of allogeneic tumor cell lysate as a DC pulsing antigen has been tested in mouse melanoma pulmonary me tastasis model. Methods: B16F10 melanoma cells $(1{\timeS}10^5/mouse)$ were inoculated intra venously into the C57BL/6 mouse. Therapeutic DCs were cultured from the bone marrow myeloid lineage cells with GM-CSF and IL-4 (1,000 U/ml each) for 7 days and pulsed with lysate of either autologous B16F10 (B-DC), allogeneic K1735 (C3H/He origin; K-DC) or CloneM3 (DBA2 origin; C-DC) melanoma cells for 18 hrs. Pulsed-DCs $(1{\times}10^6/mouse)_{[CGP1]}$ were injected i.p. twice with one week interval starting from the day 1 after tumor cell inoculation. Results: Without observable toxicity, allogeneic tumor cell lysate pulsed-DC induced the significantly better anti-tumor response (tumor scale: $2.7{\pm}0.3,\;0.7{\pm}0.3\;and\;0.3{\pm}0.2$ for saline, B-DC and C-DC treated group, respectively). Along with increased tumor specific lymphocyte proliferations, induction of IFN-${\gamma}$ secretion against both auto- and allo-tumor cell lysates was observed from the DC treated mice. (w/B16F10-lysate: $44.97{\pm}10.31,\;1787.94{\pm}131.18,\;1257.15{\pm}48.27$, w/CloneM3 lysate: 0, $1591.13{\pm}1.83,\;1460.47{\pm}86.05pg/ml$ for saline, B-DC and C-DC treated group, respectively) Natural killer cell activity was also increased in the mice treated with tumor cell lysate pulsed-DC ($8.9{\pm}_{[CGP2]}0.1,\;11.6{\pm}0.8\;and\;12.6{\pm}0.7%$ specific NK activity for saline, B-DC and C-DC treated group, respectively). Conclusion: Conclusively, promising data were obtained that allogeneic-tumor cell lysate can be used as a tumor antigen for DC-based cancer immunotherapy.

• Korean Journal of Head & Neck Oncology
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• v.13 no.1
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• pp.3-8
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• 1997

Malignant melanoma has a very poor prognosis compared to other cancers. There are no specific tumor markers other than clinical staging and depth of invasion to predict the prognosis of the malignant melanoma. The nm23 has been known to inhibit the metastasis of the malignant melanoma, some studies showed that it is highly expressed in the malignant melanoma cell line which has a relatively weak metastatic potential. In this study, we compared the expression of nm23 in mucosal type with that in cutaneous type of the malignant melanoma in the head and neck according to the stage and survival rate to identify the role of nm23 expression as a prognostic factor in mucosal melanoma of the head and neck. Six out of eight cases in mucosal type and seven out of 11 cases in cutaneous type expressed nm23, which showed no significant differences. Between the two groups there were no significant differences in expression of nm23 according to clinicopathologic staging or two year survival rate. However, in cases with low cliniopathological staging and those surviving more than two years the expression was significantly increased which suggests that expression of nm23 can be used as an aid in determining the prognosis of mucosal melanoma.

• Bulletin of the Korean Chemical Society
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• v.32 no.4
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• pp.1209-1214
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• 2011

The synthesis of the series of pyrimidinylamines 1a-d and pyrimidinylureas 1e-u bearing a novel pyridinylcarbonylpyrimidine scaffold and their antiproliferative activities against A375 human melanoma cell line were described. Among them, three compounds 1e, 1h, and 1o showed superior antiproliferative activities to Sorafenib ($IC_{50}=5.5{\mu}M$) as a reference compound. In our series, urea compound 1o having 4-chloro-3-trifluoromethyl moiety on the benzene nucleus exhibited very good antiproliferative activity with $IC_{50}$ value of $1.4{\mu}M$.

• 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.

• Archives of Pharmacal Research
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• v.29 no.3
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• pp.235-240
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• 2006

The oral administration of extracts of young radishes cultivated with sulfur after intravenous tumor cell injection achieved a marked reduction of pulmonary colonization in mice. Treatment of the mice with extracts of young radish cultivated with sulfur did not show any increase in the number of CD8+ or NK T cells in the spleen, indicating no influence on host immunity. Sulforaphane, which could be a candidate for an active compound from young radishes cultivated with sulfur, inhibited cell growth of B16-F10 melanoma cells. In addition, extracts of the young radish cultivated with sulfur-fed group showed enhanced quinine reductase (QR) activities in the liver and lung and a slight increase of glutathione S-transferase (GST) activity in the liver. These results suggested that the administration of extracts of young radishes cultivated with sulfur suppressed pulmonary tumorigenesis, possibly due to increased activity of detoxification enzymes in the liver and lung, and partly due to cell cytotoxicity.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.20 no.3
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• pp.14-28
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• 2007

Objective : Hwagi-Jokyeong-Tang (化氣調經湯, HJT) was described in DongeuiBogam(東醫寶鑑). This remedy has been used to treat patients with Naryeok, which is similar as tuberculous cervical lymphadenitis in western medicine. Methods : In this study, the present author investigated the effects of HJT on on Human HaCaT keratinocyte and malignant melanoma cells such as SK-MEL-2 and B16F10 in terms of cell viabilities, proliferations, DPPH free radical scavenging activities, oxygen free radical productions and inhibitory action on elastase activities. Results : HJT acceleated proliferation of HaCaT keratinocytes dose-dependantly. HJT also prevented cell death of HaCaT induced by Hydrogen peroxide, which products oxygen free radicals. On the contrary, HJT did not affect proliferations of SK-MEL-2 or B16F10. In addition, HJT was shown to have DPPH free radical scavenging activities and also have inhibitory effects on elastase activities too. On the fluorescent examinations, the present author know that HJT did not affect production levels of oxygen free radicals in malignant melanoma cell, SK-MEL-2. Conclusions : These results suggest that HJT has possibilities of usage for functional cosmetics which have skin regeneration or prevention from skin tissue injury.

• Journal of Microbiology and Biotechnology
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• v.29 no.8
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• pp.1204-1211
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• 2019

Fungal exopolysaccharides are important natural products having diverse biological functions. In this study, exopolysaccharides from Fomitopsis castanea mycelia (FEPS) were prepared, and the highest mushroom tyrosinase inhibitory activity was found. FEPS were prepared from cultivation broth by ethanol precipitation method. The extraction yield and protein concentration of FEPS were 213.1 mg/l and 0.03%, respectively. FEPS inhibited mushroom tyrosinase with the half maximal inhibitory concentration ($IC_{50}$) of 16.5 mg/ml and dose-dependently inhibited cellular tyrosinase activity (63.9% at $50{\mu}g/ml$, and 83.3% at $100{\mu}g/ml$) in the cell-free extract of SK-MEL-5 human melanoma cell and ${\alpha}$-melanocyte-stimulating hormone (${\alpha}-MSH$)-stimulated melanin formation in intact SK-MEL-5 human melanoma cell. The $IC_{50}$ of FEPS against NO production from RAW264.7 macrophage cells was $42.8{\pm}0.64{\mu}g/ml$. By in vivo study using a zebrafish model, exposure of FEPS at $400{\mu}g/ml$ to dechorionated zebrafish embryos for 18 h decreased the pigment density, compared to that without FEPS-treated control.