• 대한수의학회지
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• 제58권2호
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• pp.65-72
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• 2018

The present study observed the effects of a green tea (Camellia sinensis) flower extract (GTFE) on melanin synthesis in B16-F10 melanoma cells. GTFE exhibited antioxidant activity on 2,2-diphenyl-1-picrylhydrazyl and inhibited mushroom tyrosinase activity in a dose-dependent manner. Furthermore, GTFE significantly diminished ${\alpha}-melanocyte$ stimulating hormone (${\alpha}-MSH$) stimulated cellular melanin content and tyrosinase activity throughout the concentration range evaluated. Based on RNA sequencing analysis, differential gene expression patterns observed in ${\alpha}-MSH$ stimulated B16-F10 melanoma cells were normalized by the addition of GTFE. In particular, the expression levels of melanoregulin and tyrosinase genes which are key regulating genes in melanin synthesis were up-regulated by 3.5 and 3 fold respectively by ${\alpha}-MSH$, and were normalized to control levels by the addition of GTFE. The results suggest that GTFE inhibits melanin synthesis in ${\alpha}-MSH$ stimulated B16-F10 melanoma cells by normalizing expression of genes that are essential for melanin synthesis. Overall, the results suggest that GTFE could be applied in the development of a whitening agent for the treatment of dermal hyperpigmentation.

• Asian Pacific Journal of Cancer Prevention
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• 제17권5호
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• pp.2587-2592
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• 2016

Background: Early onset sporadic colorectal cancer (CRC) is a biologically and clinically distinct entity hypothesized to exhibit differences in histological features and microsatellite instability (MSI) as compared to typical onset CRC. This study compared the MSI status, mismatch repair enzyme deficiency and clinicopathological features of early onset (aged ${\leq}45$ years) with controls (>45 years). Materials and Methods: A total of 30 cases and 30 controls were analyzed for MSI status using the Bethesda marker panel. Using antibodies against hMLH1, hMSH2 and hMSH6, mismatch repair protein expression was assessed by immunohistochemistry. Molecular characteristics were correlated with clinicopathological features. Results: The early onset sporadic CRCs were significantly more poorly differentiated tumors, with higher N2 nodal involvement and greater frequency of signet ring phenotype than the typical onset cases. MSI was observed in 18/30 cases, with 12/18 designated as MSI-high (MSI-H) and 6/18 designated as MSI-low (MSI-L). In the control group, 14 patients exhibited MSI, with 7 MSI-H and 7 MSI-L. MSI tumors in both cases and controls exhibited loss of hMLH1, hMSH2 and hMSH6. MSS tumors did not exhibit loss of expression of MMR proteins, except hMLH1 protein in 3 controls. No statistically significant difference was noted in MSI status or expression of MMR proteins in cases versus controls. Conclusions: Microsatellite status is comparable between early and typical onset sporadic CRC patients in Pakistan suggesting that differences in clinicopathological features between these two subsets are attributable to other molecular mechanisms.

• Fisheries and Aquatic Sciences
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• 제22권5호
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• pp.11.1-11.8
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• 2019

Background: In the present study, the skin-whitening effects of a marine-sourced mixture that includes a fucoidanrich extract of Undaria pinnatifida (UPEF), a phlorotannin-rich extract of Ecklonia cava (ECE), and glycosaminoglycans (GAGs) from sea squirt skin were investigated. Methods: The whitening effects of the mixture and its components were evaluated by measuring the inhibition of mushroom tyrosinase and melanin synthesis in alpha-melanocyte-stimulating hormone (${\alpha}$-MSH)-stimulated B16F10 melanoma cells. Results: Each component alone markedly inhibited mushroom tyrosinase in a dose-dependent manner, and in ${\alpha}$-MSH-stimulated B16F10 cells, they inhibited melanin synthesis and were cytotoxic. However, the whitening effects of UPEF, ECE, and GAGs in combination were greater than those of each component alone. A mixture in the ratio of 4:5:1 (UEG-451) showed the strongest activity without cytotoxicity. Further study suggested that UEG-451 inhibits ${\alpha}$-MSH-stimulated melanogenesis in B16F10 cells by downregulating tyrosinase and tyrosinase-related proteins, such as TRP-1 and TRP-2, via the inhibition of MITF expression. Conclusions: These results suggest that mixing the different components at optimum ratios might be an effective way to improve their bioactivities and reduce toxicity and that UEG-451 possesses strong whitening effects that could be used in the cosmetic industry.

• 대한화장품학회지
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• 제24권3호
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• pp.65-72
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• 1998

In this study, we demonstrated the whitening effect of Lagenaria leucantha through the melanin biosynthesis of S bikiniensis and inhibition of melanogenesis in cultured Bl6 melanocytes. And we confirmed the whitening effect of Lagenaria leucantha through the depigmentation of gold fish in vivo. The melanogenesis of B$_{16}$ melanocytes was founded to be activated dose and time dependently by the treatment of u- MSH. When the B$_{16}$ melanocytes was treated with 200nM of $\alpha$-MSH, the morphology of melanocytes was remarkably changed. The melanin content and the synthesis of tyrosinase were strikingly increased. Lagenaria ieucantha inhibited the melanin formation stimulated by $\alpha$-MSH without affection of cell viability. However, Lagenaria leucantha didn't inhibit tyrosinase activity and showed weak suppression on the synthesis of tyrosinase. These results suggest Lagenaria leucantha might inhibit melanin formation with tyrosinase independent manner. Lagenaria ieucantha also inhibition melanin biosynthesis with 18mm inhibition zone in S.bikiniensis. To evaluate the inhibitory activity of melanogenesis of Lagenaria leucantha in vivo, we examined its effect on depigmentation of gold fish. Lagenaria ieucantha remarkably reduced the size and density of melanophores in gold fish. These results suggest that Lagenaria ieucantha can be used as a whitening agent in cosmetics.ics.s.

• 한국동물생명공학회지
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• 제38권3호
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• pp.99-108
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• 2023

Background: Efficient gene editing technology is needed for successful knock-in. Homologous recombination (HR) is a major double-strand break repair pathway that can be utilized for accurately inserting foreign genes into the genome. HR occurs during the S/G2 phase, and the DNA mismatch repair (MMR) pathway is inextricably linked to HR to maintain HR fidelity. This study was conducted to investigate the effect of inhibiting MMR-related genes using CdCl₂, an MMR-related gene inhibitor, on HR efficiency in HC11 cells. Methods: The mRNA and protein expression levels of MMR-related genes (Msh2, Msh3, Msh6, Mlh1, Pms2), the HR-related gene Rad51, and the NHEJ-related gene DNA Ligase IV were assessed in HC11 cells treated with 10 μM of CdCl₂ for 48 hours. In addition, HC11 cells were transfected with a CRISPR/sgRNA expression vector and a knock-in vector targeting Exon3 of the mouse-beta casein locus, and treated with 10 μM cadmium for 48 hours. The knock-in efficiency was monitored through PCR. Results: The treatment of HC11 cells with a high-dose of CdCl₂ decreased the mRNA expression of the HR-related gene Rad51 in HC11 cells. In addition, the inhibition of MMR-related genes through CdCl₂ treatment did not lead to an increase in knock-in efficiency. Conclusions: The inhibition of MMR-related gene expression through high-dose CdCl₂ treatment reduces the expression of the HR-related gene Rad51, which is active during recombination. Therefore, it was determined that CdCl₂ is an inappropriate compound for improving HR efficiency.

• 한국자원식물학회지
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• 제22권5호
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• pp.477-482
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• 2009

한국에서 주요하게 재배되는 세 개의 품종(켐벨, 청포도, 머루포도)을 대상으로 껍질과 씨 추출물의 미백효능을 검색하였다. Mouse melanoma인 B16세포에 $\alpha$-MSH와 샘플을 처리하여 melanin 생성을 측정하였다. 세 품종의 껍질은 효과가 없었으며 켐벨과 청포도의 씨 추출물은 세포독성이 매우 강하였다. 머루포도 씨 추출물은 $50{\mu}g/ml$에서 대조구에 비해 melanin 생성은 $51.6{\pm}20.5%$ 이었으며 세포 생존율은 $90.4{\pm}11.3%$ 이어서 약간의 세포 독성에도 불구 melanin 생성 억제 효과가 뚜렷하였다. 머루포도 씨 추출물은 B16세포에서 $\alpha$-MSH에 의한 tyrosinase 발현량 증가를 억제하였다. 이후 연구는 1) 머루포도 씨의 생리활성 단일물질 검색과 2) $\alpha$-MSH의 신호전달 과정에 추출물 및 생리활성 단일물질이 어떻게 영향을 미치는지 하는 점이고 현재 실험에 진행 중에 있다.

• 한국식품위생안전성학회지
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• 제30권3호
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• pp.258-264
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• 2015

본 연구는 윈터체리 추출물의 미백 활성을 검증하여 기능성 미백 소재로서의 가능성을 확인하였다. 먼저, DPPH, ABTS, FRAP를 이용한 항산화 활성 검증에서 윈터체리 추출물은 매우 높은 항산화 효과를 보였으며, 티로시나아 제의 활성에도 농도 의존적인 억제 효과를 보여주었다. 마우스 유래 B16-F10 멜라노마 세포를 이용한 멜라닌 생성에서 윈터체리 추출물이 미치는 영향을 확인한 결과, 세포 내에서 생합성 되는 멜라닌의 양이 상당히 감소하고 있음을 확인하였다. 또한, 멜라닌 생성을 더욱 유도하는 ${\alpha}-MSH$를 세포에 처리하였을 때, 윈터체리 추출물은 농도 의존적인 억제 효능을 보여주었다. 이러한 윈터체리 추출물의 멜라닌 생성 억제는 MITF 전사인자의 발현을 억제함으로써 일어나고, 결과적으로 멜라닌 생합성과 관련된 티로시나아제와 Tyrp-1 등의 단백질 발현을 감소시킴으로써 일어나게 됨을 확인하였다. 이러한 결과들로 볼 때, 윈터체리 추출물은 기존의 미백 원료들을 대체할 수 있을 뿐만 아니라 함께 사용할 경우 상승 효과를 낼 수 있는 새로운 미백 소재로 활용될 수 있을 것이다.

• 대한본초학회지
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• 제28권3호
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• pp.69-74
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• 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.

• The Korean Journal of Physiology and Pharmacology
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• 제22권1호
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• pp.53-61
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• 2018

Ethyl linoleate is an unsaturated fatty acid used in many cosmetics for its various attributes, such as antibacterial and anti-inflammatory properties and clinically proven to be an effective anti-acne agent. In this study, we investigated the effect of ethyl linoleate on the melanogenesis and the mechanism underlying its action on melanogenesis in B16F10 murine melanoma cells. Our results revealed that ethyl linoleate significantly inhibited melanin content and intracellular tyrosinase activity in ${\alpha}$-MSH-induced B16F10 cells, but it did not directly inhibit activity of mushroom tyrosinase. Ethyl linoleate inhibited the expression of microphthalmia-associated transcription factor (MITF), tyrosinase, and tyrosinase related protein 1 (TRP1) in governing melanin pigment synthesis. We observed that ethyl linoleate inhibited phosphorylation of Akt and glycogen synthase kinase $3{\beta}$ ($GSK3{\beta}$) and reduced the level of ${\beta}-catenin$, suggesting that ethyl linoleate inhibits melanogenesis through $Akt/GSK3{\beta}/{\beta}-catenin$ signal pathway. Therefore, we propose that ethyl linoleate may be useful as a safe whitening agent in cosmetic and a potential therapeutic agent for reducing skin hyperpigmentation in clinics.

• Molecules and Cells
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• 제26권6호
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• pp.576-582
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• 2008

Nervous system development takes place after positional information has been established along the dorsal-ventral (D/V) axis. The initial subdivision provided by a gradient of nuclear dorsal protein is maintained by the zygotic genes expressed along the D/V axis. In this study, an investigation was conducted to determine the range of Dpp function in repressing the expression of eagle (eg) that is present in intermediate neuroblasts defective (ind) and muscle specific homeobox (msh) gene domain. eg is expressed in neuroblast (NB) 2-4, 3-3 and 6-4 of the msh domain, and NB7-3 of the ind domain at the embryonic stage 11. In decapentaplegic (dpp) loss-of-function mutant embryos, eg was ectopically expressed in the dorsal region, while in dpp gain-of-function mutants produced by sog or sca-GAL4/UAS-dpp, eg was repressed by Dpp. It is worthy of note that Dpp produced from sim;;dpp embryos showed that Dpp could function at long range. However, Dpp produced from en-GAL4/UAS-dpp or wg-GAL4/UAS-dpp primarily acted at short-range. This result demonstrated that this discrepancy seems to be due to the repression of Dpp to EGFR signaling in sim;;dpp embryos. Taken together, these results suggest that Dpp signaling works at short-range, but can function indirectly at long-range by way of repression of EGFR signaling during embryonic neurogenesis.