• IMMUNE NETWORK
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• 제5권3호
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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.

• 한국식품영양과학회지
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• 제34권9호
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• pp.1325-1329
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• 2005

유전자발현 조절 수준에서 멜라닌 색소 생합성 억제물질을 탐색하고자 지치 뿌리로부터 유용성 물질을 추출${\cdot}$분획하여 tyrosinase 프로모터를 지닌 B16 mouse melanoma cell에 처리한 결과 그 메탄을 추출물은 $10{\mu}g/mL$의 농도에서 약 $33\%$ 이상의 tyrosinase프로모터 활성 억제효과를 나타내었으며, 세포 활성능이 $100{\mu}g/mL$의 농도에서 약 $108\%$로 매우 안전하였다. 그리고 methylene chloride, ethyl acetate, butyl alcohol, 물 등의 용매 분획물들도 농도에 따라 차이는 있었지만 $100{\mu}g/mL$또는 $500{\mu}g/mL$의 고농도에서 tyro-sinase 프로모터의 활성을 억제하였다 또한, 지치 메탄올 추출물의 농도를 달리하여 3일 동안 처리하였을 때 멜라닌색소의 생성능은 $10{\mu}g/mL$와 $100{\mu}g/mL$의 농도에서 대조군 세포에 비 해 각각 약 $11\%$와 $24\%$로 크게 감소하는 결과를 보여주었다.

• 치위생과학회지
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• 제4권2호
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• pp.81-84
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• 2004

천연물로부터 치은에 생성되는 멜라닌의 양을 효과적으로 조절하여 치은 미백을 목표로 하는 물질을 분리하고자 문헌에 소개되고 있는 미백 효과를 가진 전통한약을 비롯하여, 민간에서 사용되고 있는 약 100여 종의 고등식물을 대상으로 B-16 mouse melanoma cell lines에서 melanin 생성 억제 효과를 screening하였다. 그 결과 천궁의 methylene chloride 분획에서 활성을 나타내어 실험재료로 선택하여 2개의 화합물을 분리하였으며 각종 spectral data를 검토하여 linoleic acid methyl ester(1), 1,3-dilinoleoyl-2-stearoyl glycerol(2)로 구조를 규명하였다. 이들 화합물은 B-16 mouse melanoma cell lines에서의 melanin 생성억제 활성을 Kojic acid를 비교 물질로하여 측정하였다. 2종의 화합물 중 1,3-dilinoleoyl-2-stearoyl glycerol은 높은 활성을 보이지 않았으나 linoleic acid methyl ester는 Kojic acid에 비하여 강한 활성을 나타내었다.

• 생약학회지
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• 제35권4호통권139호
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• pp.324-329
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• 2004

SKT is consisted of skullcap radix, knope-sedge radix and trametes mushroom. SKT mixture extract has been used for curing breast cancer and cervical cancer as a folk medicine without any kind of experimental evidence to support the rationales for its clinical use. This study was undertaken to investigate the antitumor effects and toxicity of SKT. Tumor was induced by implantation of B16F10 melanoma cells $(1{\times}10^6\;cells/mouse)$ into abdominal skin in ICR mice and SKT application (5 mg/mouse, p.o.) was initiated 4 days prior to tumor induction and lasted for 42 days. SKT significantly inhibited not only tumor growth but also metastasis of i.v. implanted melanoma cells into lung and showed prolonged life span of tumor bearing mice. The combined theraphy of SKT with doxorubicin was more effective against tumor metastasis into lung. SKT almostly recovered serum SGPT to normal level of galactosamine/LPS-induced hepatitis mice. High dose of SKT did not show any acute side effects. But, in vitro SKT did not inhibit the growth of melanoma cells, which suggests that the antitumor effects of SKT might be menifested by indirect mechanisms.

• Biomolecules & Therapeutics
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• 제20권2호
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• pp.165-170
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• 2012

Cell migration plays a role in many physiological and pathological processes. Reactive oxygen species (ROS) produced in mammalian cells influence intracellular signaling processes which in turn regulate various biological activities. Here, we investigated whether melanoma cell migration could be controlled by ROS production under normoxia condition. Cell migration was measured by wound healing assay after scratching confluent monolayer of B16F10 mouse melanoma cells. Cell migration was enhanced over 12 h after scratching cells. In addition, we found that ROS production was increased by scratching cells. ERK phosphorylation was also increased by scratching cells but it was decreased by the treatment with ROS scavengers, N-acetylcysteine (NAC). Tumor cell migration was inhibited by the treatment with PD98059, ERK inhibitor, NAC or DPI, well-known ROS scavengers. Tumor cell growth as judged by succinate dehydrogenase activity was inhibited by NAC treatment. When mice were intraperitoneally administered with NAC, the intracellular ROS production was reduced in peripheral blood mononuclear cells. In addition, B16F10 tumor growth was significantly inhibited by in vivo treatment with NAC. Collectively, these findings suggest that tumor cell migration and growth could be controlled by ROS production and its downstream signaling pathways, in vitro and in vivo.

• 약학회지
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• 제51권5호
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• pp.350-354
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• 2007

Taurine has been shown to be tissue-protective against oxidant-induced injury and is a powerful regulator of the immune system. However, there is no study on the antimelanogenic effect of taurine. In this study, we investigated the whitening effect of taurine in B16F10 mouse melanoma cells. Cell viability was measured by MTT assay. We examined melanin contents and tyrosinase activity according to time and concentration. Extracellular signal regulated kinase (ERK) is an important regulator of melanogenesis. It has been reported that activated ERK induced microphthalmia associated transcription factor (MITF) phosphorylation and its subsequent degradation and thus reduced melanin synthesis. In our B16F10 cell culture system, taurine led to decrease melanin contents by 21% at 48 hr. We then observed taurine effects on ERK-P, MITF and tyrosinase by Western blot. ERK was activated at 18 hr and 24 hr, whereas MITF reduced. We could not observe any differences in the levels of tyrosinase. These results suggested that taurine inhibited melanogenesis by ERK signal pathway via MITF degradation. We expect that taurine has potential skin whitening agents in cosmetics.

• 대한화장품학회:학술대회논문집
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• 대한화장품학회 2003년도 IFSCC Conference Proceeding Book II
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• pp.241-242
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• 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.

• 한국균학회지
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• 제38권2호
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• pp.167-171
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• 2010

The present study aims to evaluate Cordyceps militaris water extract (CMWE) with a view to develop melanogenesis inhibitors. Inhibitory activities of CMWE against tyrosinase, L-DOPA(L-3,4-dihydroxyphenylalanine) oxidation, and melanin biosynthesis in B16 mouse melanoma cells were investigated. CMWE, at $5000\;{\mu}g/ml$, inhibited tyrosinase activity of 71% and DOPA oxidation of 40% as reacting with L-DOPA. Furthermore, B16 mouse melanoma cell survived over 50% from low to high dose on MTT assay, and CMWE markedly inhibited (> 50%) melanin synthesis at $5000\;{\mu}g/ml$. The inhibitory effect of CMWE on melanogenesis was attributed to enhancement of tyrosinase degradation. Key enzyme of melanin biosynthesis is tyrosinase which catalyses a beginning step from tyrosine to DOPA quinine and melanin formation step, respectively. These results indicated that CMWE may be a potential source of novel whitening agents for cosmetic or therapeutic application.

• 생약학회지
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• 제41권4호
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• pp.238-244
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• 2010

Fourteen compounds were isolated from the roots of Cynanchum auriculatum and their chemical structures were identified as ${\beta}$-sitosterol (1), acetovanillone (2), p-hydroxyacetophenone (3), 2,4-dihydroxyacetophenone (4), 2,5-dihydroxyacetophenone (5), cynandione A (6), methyleugenol (7), daucosterol (8), Succinic acid (9), cynauriculoside A (10), wilfoside C3N (11), wilfoside C1N (12), wilfoside K1N (13) and wilfoside C1G (14). Among them, compounds 2-5 were isolated from this plant for the first time. And 2, 5-dihydroxyacetophenone (5) showed the most potent inhibitory effect on melanogenesis in B-16 mouse melanoma cell lines with $IC_{50}$ value of $20\;{\mu}M$.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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• pp.137.2-137.2
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• 2003

Dendritic cells (DCs) are known to professional antigen presenting cell (APC). Due to the role as an effective activator of cytotoxic T Lymphocytes by expressing MHC, adhesion and co-stimulatory molecules, DCs are now widely recognized to play an important role in the immune responses to tumors.We investigated the effect of cultured DCs in murine melanoma pulmonary metastasis model. To follow the metastasis protocol, syngenic melanoma cells were inoculated intra-venously into the mouse (B16F10 into the C57BL/6)8 days prior to the first DC injection (1$\times$106 DCs/ mouse, i.p.) and the autologous tumor cell lysate pulsed-DCs were injected as a therapeutic module twice in two weeks. (omitted)