• Journal of Life Science
• /
• v.28 no.8
• /
• pp.900-907
• /
• 2018

This study was carried out to evaluate the possibility of unripe apple peel water extracts as cosmetic materials and to evaluate the biological activities of the antioxidant and whitening effects of the samples. The antioxidative properties of the samples were confirmed by 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) cation radical scavenging ability. To evaluate the whitening effect of the samples, several analytical techniques were used, including toxicity evaluations of the samples by MTT assays. Measurements of the inhibition rates of cellular tyrosinase, melanin synthesis rates, and expression rates of whitening-related proteins and genes were confirmed using melanoma (B16F10 cell). At equivalent unripe apple peel water concentrations ($1,000{\mu}g/ml$), the DPPH radical scavenging and the ABTS cation radical scavenging activities were 77.3% and 93.1%, respectively. The whitening activity evaluation showed that tyrosinase activity and melanin synthesis were inhibited by 19.8% and 17.3%, respectively, at unripe apple peel water extract concentrations of $50{\mu}g/ml$. In B16F10 cells induced by ${\alpha}$-MSH, the expression of tyrosinase, TRP-1, and TRP-2 decreased. Also, the activity of the transcription factor MITF was inhibited. In real-time PCR experiments, the expression of related genes at the upstream signal level was also found to be progressively lowered as the concentration of unripe apple peel water extracts increased. From these results, it was confirmed that the unripe apple peel water extracts showed excellent whitening efficacy and could be used as safe, natural, raw cosmetic material in the future.

• Environmental Mutagens and Carcinogens
• /
• v.25 no.1
• /
• pp.1-5
• /
• 2005

The present work was undertaken to evaluate the antimetastatic potential of capsaicin (8­methyl-N-vanillyl-6-nonenamide) by measuring its effects on matrix metalloproteinase activity, cell invasion and lung metastasis. Significant inhibition of matrix metalloproteinase-2 activity by capsaicin (100 $\mu$M) was detected by gelatin zymography. In vitro invasion assay showed capsaicin (50, 100 $\mu$M) reduced tumor cell invasion ($28-40\%$). Capsaicin (i.p., 2.5 mg/kg) inhibited development of lung colonization ($58\%$). These results suggest that capsaicin prevents metastasis in part through suppression of invasion of B16F10 melanoma cells by inhibiting matrix metalloproteinase-2 responsible for degradation of extracellular matrix.

• The Korea Journal of Herbology
• /
• v.29 no.6
• /
• pp.157-163
• /
• 2014

Objectives : Gamisoyo-san complex prescription were made with Angelicae Gigantis Radix, Paeoniae Radix, Atractylodes rhizome white, Hoelen, Bupleuri Radix, Moutan Cortex Radicis, Gardeniae Fructus, Zingiberis Rhizoma Crudus, Menthae Herba. The purpose of this study was to research the whitening effect of the extract from Gamisoyo-san, which is one of the used herbal complex prescription. Methods : This study investigated inhibitory effect of Gamisoyo-san in tyrosinase activity. Cell viability were performed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Then, Gamisoyo-san measured reversed-transcription-PCR for mRNA expression using B16F10 mouse melanoma cells. Results : For whitening effects, the tyrosinase inhibition effect of extract was shown to 52.4% at $5,000{\mu}g/m{\ell}$ concentration. The cell viability on B16F10 melanoma cells of Gamisoyo-san extract showed higher than 75% at $1,000{\mu}g/m{\ell}$ concentration. In this study, an experiment was performed by setting the non-toxic concentration range of 50, 150, $250{\mu}g/m{\ell}$. The Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as a positive control. The microphthalmia-associated transcription factor (MITF), tyrosinase related protein-1 (TRP-1), tyrosinase related protein-2 (TRP-2), tyrosinase mRNA expression inhibitory by reverse transcription-PCR of Gamisoyo-san extract were decreased by 95.3%, 98.8%, 96.3% and 49.5% at $250{\mu}g/m{\ell}$ which the highest concentration. Conclusions : All these findings could verify that whitening effects of Gamisoyo-san extract by tyrosinase inhibitory activity and mRNA expression. The Gamisoyo-san could be used as material for functional cosmetics, such as skin whitening products.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
• /
• v.28 no.3
• /
• pp.30-47
• /
• 2015

Objective : Melanoma is a very critical and devastating disease. Although many people have depended on surgical operation in melanoma treatment, they have placed importance on non-invasive methods constantly. So we planned to establish a research methodology by analysing existing articles containing conservative melanoma treatments in Journals of Korean Medicine published in Korea.Methods : Using search words of anti-cancer, B16, cancer, lung metastasis, melanoma, metastasis, S-100, SK-MEL, tumor, tyrosinase, we collected 26 articles by searching internet portal sites as following;Using search words of anti-cancer, B16, cancer, lung metastasis, melanoma, metastasis, S-100, SK-MEL, tumor, tyrosinase, we collected 26 articles by searching internet portal sites as followinghttp://oasis.kiom.re.kr,http://www.koreantk.com,http://www.riss.kr,http://www.dbpia.co.kr,http://www.ndsl.kr,http://kiss.kstudy.com,http://www.naver.com,http://www.google.com.Result : The number of articles is 26 and in the year of 2003, 2004 is ranked the highest number in publication. The journal of acupuncture & moxibustion society ranked the highest(30.8%). 2 and 4 authors ranked the highest(26.9%) in number of authors. T-test ranked the highest(58.1%) in statistics methods. P.O. med indicated in 11 articles and Pharmacopuncture in 15 articles. B16 murine melanoma cell was indicated in 25 articles by cancer-induced methods. In measurement, T cell activity was indicated in 14 articles, NK activity in 4 articles, IL-2 in 6 articles, apoptosis in 1 article, lung metastasis in 14 articles.Conclusion : Considering overall results, it is necessary to diversify cancer-induced methods and measurement methods in experimental melanoma research.

• Journal of the Korean Applied Science and Technology
• /
• v.33 no.3
• /
• pp.606-613
• /
• 2016

The purpose of this study is to analyze the possibility of a residual product of coffee (RC). RC oil extracted with n-hexane at $60({\pm}10)^{\circ}C$ for 24 hours. In this study, the cytotoxicity of RC oil was observed against B16F10 melanoma cells and RAW 264.7 macrophage cells by water solubletetrazolium salt-1 assay, and The RC oil measured by methods of DPPH radical scavenging and antimicrobial activities in Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Candida albicans. As a result, the RC oil treatment-related cytotoxic effects appeared on B16F10 melanoma cells from 0.125 to $2{\mu}{\ell}/m{\ell}$ and RAW 264.7 macrophage cells from 0.125 to $0.5{\mu}{\ell}/m{\ell}$ concentrations in this study. RC oil is radical scavenging activity concentrations on dependent. The antimicrobial activity of RC oil ($150{\mu}l/{\ell}$) was determined by clear zone method. Straphylococcus epidermidis, Straphylococcus aureus, Escherichia coli, Candida albicans showed clear zone by each $11.3{\pm}0.4$, $12.{\pm}0.7$, $12.0{\pm}0.0$, $0.0{\pm}0.0mm$. It is suggested that RC oil have effects on the cytotoxicity, antioxidant and antimicrobial that could be applicable to cosmetics as a new material.

• IMMUNE NETWORK
• /
• v.6 no.3
• /
• pp.154-162
• /
• 2006

Background: Dendritic cell (DC)-based cancer immunotherapy is studied for several years. However, it is mainly derived from autologous PBMC or leukapheresis from patient, which has limitations about yield and ability of DC production according to individual status. In order to solve these problems, inquiries about allogeneic DCs are performed but there are no preclinical trial answers for effect or toxicity of allogeneic DC to use for clinical trial. In this study, we compared the anti-tumor effect of allogeneic and autologous DCs from mouse bone marrow stem cells in mouse metastatic melanoma model. Methods: B16F10 melanoma cells ($5{\times}10^4$/mouse) were injected intravenously into the C57BL/6 mouse. Therapeutic DCs were differentiated from autologous (C57BL/6: CDC) or allogeneic (B6C3F1: BDC) bone marrow stem cells with GM-CSF, SCF and IL-4 for 13days and pulsed with B16F10 tumor cell lysate (Blys) for 18hrs. DC intra-peritoneal injections began on the 8th day after the tumor cell injection by twice with one week interval. Results: Anti-tumor response was observed by DC treatment without any toxicity especially in allogeneic DC treated mice (tumor burden score: $2.667{\pm}0.184,\;2.500{\pm}0.463,\;2.000{\pm}0.286,\;1.500{\pm}0.286,\;1.667 {\pm}0.297$ for saline, CDC/unpulsed-DC: U-DC, CDC/Blys-DC, BDC/U-DC and BDC/Blys-DC, respectively). IFN-${\gamma}$ secretion was significantly increased in allogeneic DC group stimulated with B16F10 cell lysate ($2,643.3{\pm}5,89.7,\;8,561.5{\pm}2,204.9.\;6,901.2{\pm}141.1pg/1{\times}10^6$ cells for saline, BDC/U-DC and BDC/Blys-DC, respectively) with increased NK cell activity. Conclusion: Conclusively, promising data was obtained that allogeneic DC can be used for DC-based cancer immunotherapy.

• The Journal of Korean Obstetrics and Gynecology
• /
• v.30 no.2
• /
• pp.49-70
• /
• 2017

Objectives: The present study is to observe the skin-regeneration, anti-wrinkle, whitening and skin moisturizing effects of Cheongsangbangpung-tang (CSBPT) with cytotoxicity. Methods: In the present study, cytotoxicity of CSBPT lyophilized aqueous extracts (yield=18.71%) was experimented against human normal fibroblast cells and B16F10 murine melanoma cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium Bromide (MTT) assay, and skin regeneration and anti-wrinkle effects were also showed through the assay of collagen type I synthesis by an enzyme immunoassay (EIA) kit as comparing with transforming growth factor (TGF)-${\beta}1$, hyaluronidase, collagenase and matrix metalloproteinase (MMP)-1 inhibitory assays as comparing with oleanolic acid (OA), and elastase inhibitory effects as comparing with phosphoramidon disodium salt (PP). In addition, whitening effects of CSBPT were observed by tyrosinase inhibitory assay and melanin formation test in B16/F10 melanoma cells as comparing with arbutin, and skin moisturizing effects were measured through mouse skin water contents test, respectively. Results: No CSBPT treatment related cytotoxic effects were demonstrated against human normal fibroblast cells and B16/F10 murine melanoma cells. CSBPT concentration-dependent increased collagen type I synthesis at human normal fibroblast cells. It also effectively suspreessed hyaluronidase, collagenase, elastase and MMP-1 activities, which were enzymes that related to declining of ECM and formation of wrinkle. CSBPT supressed B16/F10 melanoma cells's melanin productions with tyrosinase activity, which was an enzyme connected with melanin formation, and dose-dependent and significant increases of skin water contents were detected in CSBPT treated mouse skin as compared with vehicle control skins. Conclusions: CSBPT showed favorable and enough skin regeneration, anti-wrinkle, whitening and skin moisturizing effects at least in a condition of this experiment. However, more detail mechanism and in vivo skin protective efficacy studies should be conducted in future with the screening of the biological active compounds in individual herbs of Cheongsangbangpung-tang.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.20 no.5
• /
• pp.1271-1274
• /
• 2006

We investigated the effects of aqueous extracts from twelve medical herbs on ${\alpha}$-melanocyte stimulating hormone (${\alpha}$-MSH)-induced melanogenesis in B16F10 mouse melanoma cell. The cells were incubated with ${\alpha}$-MSH and aqueous extracts 5 days and 2 days and then analysed melanin amount and tyrosinase activities, respectively. Nine aqueous extract of herbs examined at 1 mg/m${\ell}$ level decreased melanin synthesis in B16F10 cell, especially in Agastache rugosa, Leonurus siviricus and Murus bombycis. The significant decrease of released extracellular melanin were also observed in treated cells with aqueous extract from Leonurus siviricus, Murus bombycis and Ledebouriella seseioides. The ${\alpha}$-MSH-induced activation of tyrosinase was inhibited in cells treated with aqueous extract from Cuscutae semen, Angelica tenuissima and Agastache rugosa. These results suggest that herbs inhibiting melanogenesis through tyrosinase activity may apply to develop whitening drugs and cosmetics.

• Korean Journal of Pharmacognosy
• /
• v.29 no.3
• /
• pp.243-247
• /
• 1998

A diverse panel of human tumor cell lines and a mouse melanoma cell line (B16-F1) were used for the cytotoxicity test of the nine sesquiterpene lactones with ${\beta}-methylene-{\gamma}-lactone$ group isolated from Hemisteptia lyrata, Chrysanthemum zawadskii and Chrysanthemum boreale. In the cell adhesion inhibitory activity test against B16-F1 mouse melanoma cell, hemistepcin B, cumambrin B, costunolide and tulipinolide were shown significant activities with $IC_{50}$ range of 2.2, 4.1, 0.9 and $0.3\;{\mu}g/ml$, respectively. In the cytotoxicity test against human tumor cells, the most active compound was costunolide having $IC_{50}$ values of below $0.3\;{\mu}g/ml$ against all the tested cell lines except for UACC62. Cumambrin A, hendelin and costunolide exhibited more strong activity against HCT15 and UO-31 cell lines than a positive control, adriamycin. All tested compounds showed an $IC_{50}$ values of below $5.0\;{\mu}g/ml$ against all the tested cell lines.

• BMB Reports
• /
• v.42 no.12
• /
• pp.806-811
• /
• 2009

Recently, various phenolic acid phenethyl ureas (PAPUs) have been synthesized from phenolic acids by Curtius rearrangement for the development of more effective anti-oxidants. In this study, we examined the anti-tumor activity and cellular mechanism of the synthetic compound (E)-1-(3,4-dihydroxyphenethyl)-3-styrylurea (PAPU1) using melanoma B16/F10 and M-3 cells. Results showed that PAPU1 inhibited the cell proliferation and viability, but did not induce cytotoxic effects on primary cultured fibroblasts. PAPU1 induced apoptotic cell death rather than necrosis in melanoma cells, a result clearly proven by the shift of cells into sub-$G_1$ phase of the cell cycle and by the substantial increase in cells positively stained with TUNEL or Annexin V. Collectively, this study revealed that PAPU1 induced apoptosis in a caspase-dependent manner, suggesting a potential role as a cancer chemopreventive agent for melanoma cells.