• Biomolecules & Therapeutics
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• 제29권4호
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• pp.445-451
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• 2021

Salicylamide, a non-steroidal anti-inflammatory drug (NSAID), is used as an analgesic and antipyretic agent. We have previously shown that several NSAIDs have anti-melanogenic properties in B16F1 melanoma cells. In contrast, we have found that salicylamide enhances melanin contents in B16F1 melanoma cells; however, the underlying mechanism is not known. Therefore, we investigated the mechanism through which salicylamide stimulates melanogenesis. Interestingly, salicylamide enhanced diphenolase activity in a cell-free assay. Western blotting and real-time RT-PCR revealed that salicylamide increased tyrosinase expression via transcriptional activation of the Mitf gene. Together, our results indicate that salicylamide could be used as an anti-hypopigmentation agent for skin and/or hair.

• Asian Pacific Journal of Cancer Prevention
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• 제13권7호
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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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• 제63권1호
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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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• 제15권8호
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• pp.3659-3665
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• 2014

To assess inhibition mechanisms of a Phellinus igniarius (PI) extract on cancer, C57BL/6 mice were orally treated with PI extractive after or before implanting H22 (hepatocellular carcinoma ) or B16 (melanoma) cells. Mice were orally gavaged with different doses of PI for 36 days 24h after introduction of H22 or B16 cells. Mice in another group were orally treated as above daily for 42 days and implanted with H22 cells on day 7. Then the T lymphocyte, antibody, cytokine, LAK, NK cell activity in spleen, tumor cell apoptosis status and tumor inhibition in related organs, as well as the expression of iNOS and PCNA in tumor tissue were examined. The PI extract could improve animal immunity as well as inhibit cancer cell growth and metastasis with a dose-response relationship. Notably, PI's regulation with the two kinds of tumor appeared to occur in different ways, since the antibody profile and tumor metastasis demonstrated variation between animals implanted with hepatocellular carcinoma and melanoma cells.

• 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.

• Biomolecules & Therapeutics
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• 제25권2호
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• pp.213-221
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• 2017

Baicalein, a natural flavonoid obtained from the rhizome of Scutellaria baicalensis Georgi, has been reported to have anticancer activities in several human cancer cell lines. However, its antimetastatic effects and associated mechanisms in melanoma cells have not been extensively studied. The current study examined the effects of baicalein on cell motility and anti-invasive activity using mouse melanoma B16F10 cells. Within the noncytotoxic concentration range, baicalein significantly inhibited the cell motility and invasiveness of B16F10 cells in a concentration-dependent manner. Baicalein also reduced the activity and expression of matrix metalloproteinase (MMP)-2 and -9; however, the levels of tissue inhibitor of metalloproteinase-1 and -2 were concomitantly increased. The inhibitory effects of baicalein on cell motility and invasiveness were found to be associated with its tightening of tight junction (TJ), which was demonstrated by an increase in transepithelial electrical resistance and downregulation of the claudin family of proteins. Additionally, treatment with baicalein markedly reduced the expression levels of lipopolysaccharide-induced phosphorylated Akt and the invasive activity in B16F10 cells. Taken together, these results suggest that baicalein inhibits B16F10 melanoma cell migration and invasion by reducing the expression of MMPs and tightening TJ through the suppression of claudin expression, possibly in association with a suppression of the phosphoinositide 3-kinase/Akt signaling pathway.

• 동의생리병리학회지
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• 제19권1호
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• pp.196-203
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• 2005

Gunggui-tang has been used for the therapy of blood disorders in Hangbang medicine for long time. Also, Glycyrrhiza uralensis has been used for deficientblood patterns with an irregular pulse or palpitations, coughing and wheezing, and heat or cold in the lungs. Melanogenesis is a physiological process resulting in the synthesis of melanin pigments. We investigated whether the water extract of Gunggui-tang plus G. uralensis inhibited melanogenesis in B16 melanoma cells. Because the molecular events connecting the regulation in tyrosinase activity remain to be elucidated, we also aimed to determine whether Gunggui-tang gamibang(GTG) affects tyrosinase at the gene activation level in the cells. First, we showed that GTG inhibited the tyrosinase promoter activity and further, down-regulated the tyrosinase protein activity in ${\alpha}-melanocyte-stimulating$ hormone $({\alpha}-MSH)-treated$ B16 melanoma cells. GTG also resulted in a decrease of melanin content in MSH-induced melanogenesis, indicating that GTG may be a useful drug in studying the regulation of melanogenesis. The pretreatment of GTG significantly prevented phosphotransferase activity of c-Jun N-terminal kinase (JNK1) and transcriptional activation of activating protein-1 (AP-1) in MSH-treated B16 melanoma cells. These findings indicate that GTG inhibits melanogenesis of B16 melanoma cells via suppression of phosphotransferase activity of JNK1 and transcriptional activation of AP-1.

• Journal of Applied Biological Chemistry
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• 제61권3호
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• pp.267-273
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• 2018

기능성 천연물 소재로서의 가능성을 검토하고자 캐모마일 추출물의 미백 효과를 조사하고 melanin 생성 반응에 관여하는 물질 억제에 대한 기전을 규명하고자 하였다. 캐모마일을 water와 60% ethanol을 이용하여 추출하였고, 얻어진 추출물을 phenolic 농도별로 설정하여 tyrosinase 저해 활성을 알아보았을 때, water 추출물의 경우 효과가 미비하였고, 60% ethanol 추출물에서는 농도 의존적으로 저해 활성이 나타내어 melanin 생성 저해 효과가 있을 것으로 판단되었다. 캐모마일 60% ethanol 추출물을 동결건조하여 얻어진 분말을 이용하여 B16F10 melanoma cell에 대한 세포 독성을 측정 결과, $75{\mu}g/mL$의 농도에서부터 독성이 관찰되어 농도 구간을 10, 25, $50{\mu}g/mL$으로 선정하였다. ${\alpha}-MSH$로 자극한 B16F10 melanoma cell에서 melanin 생성량을 측정하여 캐모마일의 melanin 성성 억제 효능과 melanin 생성에 영향을 미치는 단백질인 tyrosinase, MITF, TRP-1, TRP-2의 단백질 발현 억제 효과를 알아본 결과, 캐모마일의 농도가 높아짐에 따라 농도 의존적으로 melanin 생성 함량이 감소하였고, tyrosinase, TRP-1, TRP-2, 등의 단백질 발현량 또한 감소하는 것을 확인할 수 있었다. 따라서 캐모마일 추출물은 B16F10 melanoma cell에서 melanin의 생성을 억제하고, melanin 생성 관련 단백질발현을 억제하는 효과가 있음을 확인하였다. 위의 결과들로 인하여 캐모마일은 미백 기능성 식품 산업화를 위한 유용한 자원으로 활용 될 것으로 예상되며, 추후 산업적 응용을 위한 지속적인 연구가 진행되어야 할 것으로 판단되었다.

• 약학회지
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• 제43권4호
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• pp.494-501
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• 1999

The skin whitening effects of pine needle extract, hop extract and chestnut inner shell extract were evaluated both in vitro and in B 16 mouse melanoma cell lines. Each extracts significantly inhibited tyrosinase activity, dopa auto-oxidation and melanin biosynthesis in vitro and in B 16 cell lines. In vitro, hop extract inhibited melanin biosynthesis 15 times stronger than kojic acid at $10{\;}\mu\textrm{g}/ml$ concentration. Each extracts were stronger inhibitors of melanin biosynthesis than kojic acid in B 16 mouse melanoma cell at less than $4{\;}\mu\textrm{g}/ml$ concentration. These results show that extracts fo pine needle, hop and chestnut inner shell could be developed as skin whitening component of cosmetics.

• 대한약학회:학술대회논문집
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• 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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• pp.318.2-318.2
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• 2002

The effects of oxidative stress on the alterations of different antioxidant enzyme activity on mouse melanoma cells(B16F10) was investigated. Oxidative stress was induced by the exposeure to hydrogen peroxide(H2O2). B 16F 10 cells were exposed Phellinus linteus Ex. in combination with H2O2 and measured the time course of changes in cell viability and antioxidant enzyme activity. CAT activity peaked at 12 hr. On the contrary, SOD and GPX activity was maximum at 6 hr. The cell viability of Pheltinus linteus extracts in combination with hydrogen peroxide was higher than hydrogen peroxide alone.