• The Korean Journal of Physiology and Pharmacology
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• v.23 no.6
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• pp.501-508
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• 2019

Rosmarinic acid (RA) is a natural polyphenolic compound that exists in many medicinal species of Boraginaceae and Lamiaceae. The previous studies have revealed that RA had therapeutic effects on hepatocellular carcinoma (HCC) in the H22-xenograft models by inhibiting the inflammatory cytokines and $NF-{\kappa}B$ p65 pathway in the tumor microenvironment. However, its molecular mechanisms of immunoregulation and pro-apoptotic effect in HCC have not been fully explored. In the present study, RA at 75, 150, and 300 mg/kg was given to H22 tumor-bearing mice via gavage once a day for 10 days. The results showed that RA can effectively inhibit the tumor growth through regulating the ratio of $CD4^+/CD8^+$ and the secretion of interleukin (IL)-2 and interferon-${\gamma}$, inhibiting the expressions of IL-6, IL-10 and signal transducer and activator of transcription 3, thereby up-regulating Bax and Caspase-3 and down-regulating Bcl-2. The underlying mechanisms involved regulation of immune response and induction of HCC cell apoptosis. These results may provide a more comprehensive perspective to clarify the anti-tumor mechanism of RA in HCC.

• Biomolecules & Therapeutics
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• v.27 no.1
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• pp.41-47
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• 2019

The apoptotic effects of shikonin (5,8-dihydroxy-2-[(1R)-1-hydroxy-4-methylpent-3-enyl]naphthalene-1,4-dione) on the human colon cancer cell line SNU-407 were investigated in this study. Shikonin showed dose-dependent cytotoxic activity against SNU-407 cells, with an estimated $IC_{50}$ value of $3{\mu}M$ after 48 h of treatment. Shikonin induced apoptosis, as evidenced by apoptotic body formation, sub-G_1$ phase cells, and DNA fragmentation. Shikonin induced apoptotic cell death by activating mitogen-activated protein kinase family members, and the apoptotic process was mediated by the activation of endoplasmic reticulum (ER) stress, leading to activation of the $PERK/elF2{\alpha}/CHOP$ apoptotic pathway, and mitochondrial $Ca^{2+}$ accumulation. Shikonin increased mitochondrial membrane depolarization and altered the levels of apoptosis-related proteins, with a decrease in B cell lymphoma (Bcl)-2 and an increase in Bcl-2-associated X protein, and subsequently, increased expression of cleaved forms of caspase-9 and -3. Taken together, we suggest that these mechanisms, including MAPK signaling and the ER- and mitochondria-mediated pathways, may underlie shikonin-induced apoptosis related to its anticancer effect.

• Journal of Pharmacopuncture
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• v.21 no.4
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• pp.268-276
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• 2018

Objectives: The purpose of this study is to investigate the anti-cancer effects of different fractions of Astragalus membranaceus (AM) in human non-small cell lung cancer (NSCLC) cells. Methods: We isolated hexane, ethyl acetate, and butanol fractions from crude ethanol extract of AM. The cell death was examined by MTT assay and trypan blue exclusion assay. Apoptosis was detected by DAPI staining, annexin V-PI double staining and cell cycle analysis. The expression of apoptosis-related proteins and mitogen-activated protein kinases (MAPKs) was examined by western blot. Results: Among various fractions of AM, the ethyl acetate fraction of AM (EAM) showed the strongest cytotoxic effect in NSCLC cells. EAM reduced the cell proliferation in a time- and dose-dependent manner in NSCLC cells. In addition, EAM induced the chromatin condensation, and increased the population of sub-G1 phase and annexin V-positive cells in a time-dependent manner, indicating that EAM induced apoptosis in NSCLC cells. Consistently, EAM enhanced the expression of cleaved caspase-8 and -9, and induced the accumulation of cleaved- poly (ADP-ribose) polymerase (PARP). Among MAPK proteins, only ERK was dephosphorylated by EAM, suggesting that ERK might be related with EAM-induced apoptosis. Conclusion: Our results clearly demonstrate that EAM exhibited anti-cancer effects in NSCLC cells by induction of apoptosis. We provide a valuable evidence which suggests that AM could be a desirable therapeutic option for treatment of NSCLC.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2019.04a
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• pp.123-123
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• 2019

Xanthone is a kind of polyphenolic compounds that contain a distinctive chemical structure with a tricyclic aromatic ring found in a few higher plant families e.g. gentian root. This compound had a variety of biological activity, for instance antioxidant, antibacterial, anti-inflammatory, and anticancer effects. However, the effect of xanthone on mast cell-mediated allergic inflammation and its associated mechanism have not been elucidated. Therefore, the aim of this study was to elucidate the anti-allergic inflammatory effects and the underlying molecular mechanism of xanthone in PMACI-stimulated human mast cells-1 (HMC-1). In this result, xanthone treatment decreased the production of histamine, pro-inflammatory cytokines such as tumor necrosis factor-a (TNF-${\alpha}$), IL-6, and IL-8 and expressions of TSLP in PMACI-stimulated HMC-cells. In addition, xanthone significantly suppressed the phosphorylation of MAPKs and the activation of NF-${\kappa}B$ signal pathway in activated mast cells. Furthermore, xanthone inhibited the activation of caspase-1, an IL-$1{\beta}$ converting enzyme, in PMACI-stimulated HMC-1 cells. These findings provide evidence that xanthone could be a potential therapeutic agent for allergy-related inflammatory disorders.

• The Korean Journal of Physiology and Pharmacology
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• v.26 no.5
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• pp.377-387
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• 2022

Benzimidazole anthelmintic agents have been recently repurposed to overcome cancers resistant to conventional therapies. To evaluate the anti-cancer effects of benzimidazole on resistant cells, various cell death pathways were investigated in 5-fluorouracil-resistant colorectal cancer cells. The viability of wild-type and 5-fluorouracil-resistant SNU-C5 colorectal cancer cells was assayed, followed by Western blotting. Flow cytometry assays for cell death and cell cycle was also performed to analyze the anti-cancer effects of benzimidazole. When compared with albendazole, fenbendazole showed higher susceptibility to 5-fluorouracil-resistant SNU-C5 cells and was used in subsequent experiments. Flow cytometry revealed that fenbendazole significantly induces apoptosis as well as cell cycle arrest at G2/M phase on both cells. When compared with wild-type SNU-C5 cells, 5-fluorouracil-resistant SNU-C5 cells showed reduced autophagy, increased ferroptosis and ferroptosis-augmented apoptosis, and less activation of caspase-8 and p53. These results suggest that fenbendazole may be a potential alternative treatment in 5-fluorouracil-resistant cancer cells, and the anticancer activity of fenbendazole does not require p53 in 5-fluorouracil-resistant SNU-C5 cells.

• BMB Reports
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• v.56 no.8
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• pp.451-456
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• 2023

Deubiquitinases (DUBs) are an essential component of the ubiquitin-proteasome system (UPS). They trim ubiquitin from substrate proteins, thereby preventing them from degradation, and modulate different cellular processes. Ubiquitin-specific protease 14 (USP14) is a DUB that has mainly been studied for its role in tumorigenesis in several cancers. In the present study, we found that the protein levels of USP14 were remarkably higher in gastric cancer tissues than in the adjacent normal tissues. We also demonstrated that the inhibition of USP14 activity using IU1 (an USP14 inhibitor) or the inhibition of USP14 expression using USP14-specific siRNA markedly reduced the viability of gastric cancer cells and suppressed their migratory and invasive abilities. The reduction in gastric cancer cell proliferation due to the inhibition of USP14 activity was a result of the increase in the degree of apoptosis, as evidenced by the increased expression levels of cleaved caspase-3 and cleaved PARP. Furthermore, an experiment using the USP14 inhibitor IU1 revealed that the inhibition of USP14 activity suppressed 5-fluorouracil (5-FU) resistance in GC cells. Collectively, these findings indicate that USP14 plays critical roles in gastric cancer progression and suggest its potential to serve as a novel therapeutic target for gastric cancer treatment.

• Journal of Acupuncture Research
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• v.25 no.2
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• pp.59-74
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• 2008

목적 : 이 연구는 봉약침의 봉독과 그 주요성분인 멜리틴이 $NF-{\kappa}B$의 활성억제와 세포자멸사 관련 단백질의 발현 조절을 통하여 세포자멸사를 유도하고 전립선 암세포주인 LNCaP 세포의 성장을 억제하는지를 확인하고 해당 기전을 살펴보고자 하였다. 방법 : 봉독이나 멜리틴을 처리한 후 LNCaP의 성장억제를 관찰하기 위해 WST-1 assay, CCK-8 assay를 시행하였고, 세포자멸사의 관찰에는 DAPI, TUNEL staining assay를 시행하였으며, 세포자멸사 조절단백질의 변동 관찰에는 western blot analysis를 시행하였고, 세포자멸사와 연관된 $NF-{\kappa}B$의 활성 변화를 관찰하기 위해 EMSA를 시행하였으며, LNCaP에서 봉독이나 멜리틴과 $NF-{\kappa}B$의 상호작용을 관찰하기 위해 transient transfection assay를 시행 시 세포생존율과 $NF-{\kappa}B$의 활성 변동을 측정하였다. 결과 : LNCaP 세포에 봉독이나 멜리틴을 처리한 후, 전립선암세포의 성장, 세포자멸사의 유발, 세포자멸사 관련 단백질의 발현, $NF-{\kappa}B$의 활성, $NF-{\kappa}B$의 p50 치환 후 $NF-{\kappa}B$의 활성과 LNCaP 세포 증식에 미치는 영향을 관찰하여 다음과 같은 결과를 얻었다. 1. LNCaP 세포에서 봉독이나 멜리틴을 처리한 후 세포자멸사가 유도되어 세포성장이 억제되었고, 세포자멸사 관련 단백질 중 분리된 PARP, caspase-9, Bax는 유의한 증가를, Bcl-2, p-Akt, MMP 13, XIAP, cXIAP는 유의한 감소를 나타내었다. 2. LNCaP 세포에서 봉독이나 멜리틴을 처리한 후 $NF-{\kappa}B$의 활성의 유의한 감소를 나타내었다. 3. LNCaP 세포에서 $NF-{\kappa}B$ p50를 치환하여 작용기를 없애고 봉독이나 멜리틴을 처리하였을 경우에도 $NF-{\kappa}B$의 활성의 유의한 감소를 나타내었다. 결론 : 이상의 결과는 봉독이나 멜리틴이 $NF-{\kappa}B$의 활성 억제를 통하여 인간 전립선암세포주인 LNCaP의 세포자멸사를 유발함으로써 증식억제 효과가 있음을 입증한 것으로 전립선암의 예방과 치료에 대한 효과적인 치료제 개발에 도움이 될 것으로 기대된다. 다만 그 기전에서 봉독이나 멜리틴은 기존연구와 달리 $NF-{\kappa}B$ p50의 작용기와 직접적으로 상호작용을 하지는 않는 것으로 보이므로 심화 연구를 요한다.

• The Korean Journal of Physiology and Pharmacology
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• v.9 no.4
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• pp.231-238
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• 2005

Neuronal apoptotic events, which result in cell death, are occurred in hypoxic/ischemic conditions. Estradiol is a female sex hormone with steroid structure known to provide neuroprotection through multiple mechanisms in the central nervous system. This study was aimed to investigate the signal transduction pathway of $CoCl_2$-induced neuronal cell death and the inhibitory effects of estradiol. Administration of $CoCl_2$ decreased cell viability in both a dose- and time-dependent manner in PC12 cells. $CoCl_2$-induced cell death produced genomic DNA fragmentation and morphologic changes such as cell shrinkage and condensed nuclei. It was found that $CoCl_2$-treated cells increased the reactive oxygen species (ROS) as well as caspase-8, -9 and -3 activities. However, pretreatment with estradiol before exposure to $CoCl_2$ prevented the reduction in cell viability reduction and attenuated DNA fragmentation and morphologic changes caused by $CoCl_2$. Furthermore, the $CoCl_2$-induced increases of ROS levels and caspases activities were attenuated by estradiol. Gene expression analysis revealed that estradiol blocked the underexpression of the Bcl-2 and ameliorated the increase in the release of cytochrome c from mitochondria into cytoplasm and Fas-ligand (Fas-L) upregulated by $CoCl_2$. These results suggest that $CoCl_2$ induce apoptosis in PC12 cells through both mitochondria- and death receptor-mediated cell death pathway. Estradiol was found to have a neuroprotective effect against $CoCl_2$-induced apoptosis through the inhibition of ROS production and by modulating apoptotic effectors associated with the mitochondria- and death-dependent pathway in PC12 cells.

• The Korea Journal of Herbology
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• v.22 no.3
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• pp.27-37
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• 2007

Objectives: Hepatocellular carcinoma is the most common primary malignant tumor of the liver worldwide. In-Jin-Ho-Tang(IJHT) has been used as a traditional Chinese herbal medicine since ancient time. and today it is widely applied as a medication for jaundice which is associated with inflammation in liver. In this study, I investigated whether methanol extract of IJHT induced HepG2 cancer cell death. Methods: Cytotoxic activity of IJHT on HepG2 cells was using XTT assay. Apoptosis induction by Ros A in HCT116 cells was verified by the induction of cleavage of poly ADP-ribose polymerase (PARP). and activation of caspase-3, -8 and -9. The release of cytochrome c from mitochondria to cytosol. the level of Bcl-2 and Bax and the expression of p53 and p21 were examined by western blotting analysis. Furthermore, MAPKs activation was analyzed by western blotting analysis. Results: IJHT induced apoptosis in HepG2 cells. And treatment of IJHT resulted in the release of cytochrome c into cytosol, decreased anti-apoptotic Bcl-2, and increased pri-apoptotic Bax expression. IJHT markedly inactivated extracellular signal-regulated kinase (ERK1/2), and activated p38 mitogen-activated protein (MAP) kinase. Sodium orthovanadate (SOV), a phosphatase inhibitor, to reverse IJHT-induced ERK1/2 inactivation and SB203580, a specific p38 MAP Kinase inhibitor efficiently blocked apoptosis of HepG2. Thus, IJHT induces apoptosis in HepG2 cells via MAP kinase modulation. Conclusion: These results indicated that IJHT has some potential for use as an anti-cancer agent.

• Korean Journal of Food Science and Technology
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• v.52 no.5
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• pp.467-475
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• 2020

Ultraviolet radiation is a major cause of photoaging, characterized by wrinkle formation and skin dehydration. Mojaban (Sargassum fulvellum), belonging to the brown algae of the Sargassaceae family, exerts anti-inflammatory, antioxidant, and anti-atopic activities. Particularly, Mojaban extract alleviated ultraviolet B (UVB)-induced inflammatory responses in human keratinocytes and BALB/c mice. Here, the dorsal skin of hairless mice was exposed to UVB for photoaging. The mice were administrated 300 mg/kg Mojaban extract (S. fulvellum extract; SFE) daily. Phenotypically, SFE reduced wrinkle formation and epidermal thickness and increased skin elasticity. SFE increased hydroxyproline content by upregulating the expression of collagen-related genes and downregulating matrix metalloproteinase expression. Furthermore, SFE increased skin moisture content and inhibited transepidermal water loss. SFE upregulated the expression of involucrin, loricrin, and transglutaminase for cornified envelope formation as well as the expression of filaggrin and caspase-14 for natural moisturizing factors. Collectively, SFE may be employed as a nutraceutical ingredient for anti-photoaging.