• Biomolecules & Therapeutics
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• 제11권2호
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• pp.112-118
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• 2003

1-Methylated $\beta$-carbolines (harmaline and harmalol) and antioxidants (N-acetylcysteine and ascorbate) reduced the loss of cell viability in differentiated PC 12 cells treated with 5 mM glutamate. $\beta$-Carbolines prevented the glutamate-induced decrease in mitochondrial membrane potential, cytochrome c release and caspase-3 activation in PC 12 cells. $\beta$-Carbolines reduced the formation of reactive oxygen species and depletion of glutathione due to glutamate in PC12 cells. $\beta$-Carbolines revealed a scavenging action on hydrogen peroxide and reduced the iron and EDTA-mediated degradation of 2-deoxy-D-ribose. The results suggest that I-methylated $\beta$-carbolines attenuate the cytotoxic effect of glutamate on PC12 cells by reducing the alteration of mitochondrial membrane permeability that seems to be mediated by oxidative stress.

• Asian Pacific Journal of Cancer Prevention
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• 제17권11호
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• pp.4893-4898
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• 2016

Objective: To investigate the impact of a Croton tiglium extract on cellular proliferation and apoptosis in a non-small cell lung cancer cell line (A549) in vitro. Methods: A Croton tiglium seed methanol extract was prepare and assessed for effects on A549 cells regarding cellular proliferation, apoptotic rates, and expression of apoptosis related genes and proteins using real-time PCR and immunofluorescence. Results: The tested Croton tiglium extract inhibited A549 cell proliferation in a dose- and time-dependent manner, with significant elevation of apoptotic indexes at various concentrations after 24 h. In addition, rates in both early and late stages were higher in treated than untreated groups, the $100{\mu}g/ml$ dose causing the highest levels of apoptosis. RT-PCR showed that A549 cells treated with $100{\mu}g/ml$ Croton tiglium extract for 24 h has markedly higher Bax mRNA expression levels and obviously lower Bcl-2 expression levels than controls, equivalent results being observed for proteins by immunofluorescence. However, the mRNA expression levels of Fas and caspase-8 were not significantly altered. Conclusion: A Croton tiglium extract can inhibit proliferation of A549 cells and promote apoptosis though Bax/Bcl-2 pathways.

• 한국환경과학회지
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• 제28권11호
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• pp.993-1004
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• 2019

Chrysanthemum boreale Makino (C. boreale) is widely distributed in Asian countries, and has traditionally been used to treat various inflammatory diseases including bronchitis. In this study, we aimed to isolate biologically active compounds from leaves and stems of C. boreale. Chemical components were purified by column chromatograpy and recyclic HPLC, and characterized from their spectral data (IR, MS, NMR). Biological activity experiments were conducted for Farnesyl-protein transferase (FPTase) activity, apoptosis and nitirc oxide (NO) release. As a results, three sesquiterpene lactones were isolated. Compound 1 (4-methoxy-8-O-acetyl-10-hydroxy-2,11(13)-guaiadiene-12,6-olide) showed strong cytotoxic activities having an average growth inhibition of 50% ($GI_{50}$) value of $1.89{\mu}g/m{\ell}$ against human colon adenocarcinoma cells. Compound 1 also showed a low half maximal inhibitory concentration ($IC_{50}$) value of $10{\mu}g/m{\ell}$ for NO release. In the caspase 3 activity, compound 1 and compound 2 (8-O-(2-carbonyl-2-butyl)-3,10-dihydroxy-4,11(13) -guaiadiene-12,6-olide) exhibited 94% and 90% apoptosis inhibition activity, respectively. Compound 3 (4,8-O-diacetyl -10-hydroxy-2(3),11(13)-guaiadiene-12,6-olide) showed a strong inhibitory effect on FPTase activity with 90% inhibitory activity at a concentration of $100{\mu}g/m{\ell}$. These results clearly show the presence of lactone compounds in the leaves and stems, which may partially contribute to the pharmacological activity of C. boreale.

• 생명과학회지
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• 제25권11호
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• pp.1331-1337
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• 2015

S-allylcysteine (SAC)은 숙성된 마늘로부터 유래된 수용성 유기황화합물로서, 여러 유형의 암세포에 대한 항암효과를 갖는 것으로 제시되어왔다. 본 논문은 in vitro 및 in vivo 연구결과에 기초하여 SAC가 세포증식, 세포사멸, 세포주기 및 전이에 미치는 세포신호전달경로와 분자적 메커니즘을 정리하였다. SAC는 Bax와 caspase-3을 포함하는 세포사멸촉진 단백질을 활성화하고 Bcl-2 세포사멸억제 단백질군을 억제하여 미토콘드리아-매개 내인성 경로를 통한 세포사멸을 초래 한다. SAC는 또한 PI3K/Akt/mTOR 및 MAPK/ERK 신호전달경로를 억제하여 NF-κB, cyclins, Cdks, PCNA 및 c-Jun의 발현과 활성을 감소시키고, 세포주기 억제단백질인 p16 및 p21의 발현을 증가시킴으로써 세포주기 억제를 유도하여 세포증식을 억제한다. 뿐만 아니라, SAC는 glutathione-s-transferase (GST)와 같은 항산화효소의 활성을 유도하여 독성물질에 의해 유도된 발암작용을 방지한다. 그리고, SAC는 MAPK/ERK 및 PI3K/Akt/mTOR/NF-κB 신호경로의 억제를 통한 전사억제조절인자 Id-1 및 SLUG의 발현억제를 통하여 초래된 COX-2의 발현감소와 E-cadherin의 발현증가에 의해 신생혈관생성과 MET의 억제를 유도함으로써 암세포의 침투와 전이를 억제한다. 따라서, SAC는 암의 예방과 치료를 위한 하나의 잠재적 화학요법제로 간주될 수 있다.

• Nutrition Research and Practice
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• 제11권2호
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• pp.105-113
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• 2017

BACKGROUND/OBJECTIVES: A high-fat diet (HFD) induces obesity, which is a major risk factor for cardiovascular disease and cancer, while a calorie-restricted diet can extend life span by reducing the risk of these diseases. It is known that health effects of diet are partially conveyed through epigenetic mechanism including DNA methylation. In this study, we investigated the genome-wide hepatic DNA methylation to identify the epigenetic effects of HFD-induced obesity. MATERIALS AND METHODS: Seven-week-old male C57BL/6 mice were fed control diet (CD), calorie-restricted control diet (CRCD), or HFD for 16 weeks (after one week of acclimation to the control diet). Food intake, body weight, and liver weight were measured. Hepatic triacylglycerol and cholesterol levels were determined using enzymatic colorimetric methods. Changes in genome-wide DNA methylation were determined by a DNA methylation microarray method combined with methylated DNA immunoprecipitation. The level of transcription of individual genes was measured by real-time PCR. RESULTS: The DNA methylation statuses of genes in biological networks related to lipid metabolism and hepatic steatosis were influenced by HFD-induced obesity. In HFD group, a proinflammatory Casp1 (Caspase 1) gene had hypomethylated CpG sites at the 1.5-kb upstream region of its transcription start site (TSS), and its mRNA level was higher compared with that in CD group. Additionally, an energy metabolism-associated gene Ndufb9 (NADH dehydrogenase 1 beta subcomplex 9) in HFD group had hypermethylated CpG sites at the 2.6-kb downstream region of its TSS, and its mRNA level was lower compared with that in CRCD group. CONCLUSIONS: HFD alters DNA methylation profiles in genes associated with liver lipid metabolism and hepatic steatosis. The methylation statuses of Casp1 and Ndufb9 were particularly influenced by the HFD. The expression of these genes in HFD differed significantly compared with CD and CRCD, respectively, suggesting that the expressions of Casp1 and Ndufb9 in liver were regulated by their methylation statuses.

• 생명과학회지
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• 제22권11호
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• pp.1571-1586
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• 2012

지구상에는 약 5,400여 종의 포유동물이 있고 그 중 약 1,000여 종은 풀을 뜯어 먹고 사는 초식동물이다. 초식동물 중에서 약 250여 종이 반추동물로 알려져 있다. 반추동물인 소와 양은 반추위에서 주로 발효가 일어나지만 비반추동물인 돼지와 사람은 맹장과 결장, 직장에서 주로 발효가 일어난다. 반추위 미생물의 종류와 우점도 Bacteroidetes 51%, Firmicutes 43% 존재하며, 사람의 대장미생물의 우점도Firmicutes 65%, Bacteroidetes 25%로 존재한다. 풀의 세포벽 구성성분은 미생물에 의해 분해, 발효에 의해 SCFA (short chain fatty acid)를 생성하게 되고 acetate, propionate, butyrate 생성비율은 60:25:15이다. 장내 primary butyrate transporter인 MCT1(monocarboxylatetransports-1)에 의해서 흡수된 butyrate는 SCFA receptor GPR43과 GPR41을 활성화시킨다. Butyrate는 강력한 anti-tumorigenic 기능을 가지고 있다. Butyrate는 다양한 cancer cell에 효과를 나타내며 세포내의 기능 조절에 기여하고, 암세포사멸을 유도하는 특성이 있다. Butyrate는 caspase의 활성화, HDAC (histone deacetylase) 활성을 억제하여apoptosis를 유도하고, p53 발현증가로 cell cycle arrest와 apoptosis를 유도한다. SCFA의 항 염증작용으로는 장 상피세포에서 IL-8 발현 감소, NO합성과 NF-${\kappa}B$ (nuclear factor ${\kappa}B$)의 활성을 억제하여 염증으로 인한 암 발생을 억제한다. Butyrate는 장 점막의 생리적 기능을 유지하는데 중요한 역할을 하며 IBD (inflammatory bowel disease) 치료법으로 이용되고 있다.

• Journal of Oral Medicine and Pain
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• 제36권1호
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• pp.11-20
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• 2011

Valproic acid(VPA)는 아주 잘 알려진 항경련제로서, 30년 동안 간질치료제로서 사용되어져 왔다. VPA는 1997년에 최초로 항암제의 효능이 밝혀졌으며, VPA의 항암효과는 히스톤탈아세틸화효소 억제제의 기전에 기인한다고 규명되었다. 17AAG(17-Allyamnio-17-demethoxygeldanamycin)는 HSP90의 억제제이며, HSP90은 세포증식과 세포생존에 관여하며, 최근 17AAG가 세포자멸사를 유도한다는 연구들이 보고되어지고 있다. 본 연구는 히스톤탈아세틸화효소억제제인 VPA와 HSP90 억제제인 17AAG의 병용처리가 사람골육종세포에 상승 세포자멸사 효과가 있는지를 알기 위해서 수행되었다. VPA과 17AAG의 병용처리가 단독처리에 비해서 효과적인 세포생존율 감소가 있는지 확인하기 위해서 trypan-blue법을 시행하였고, 세포자멸사의 유도와 증가를 확인하기 위해서 Hoechst 염색법, flow cytometry(DNA hypoploidy와 MMP 측정), Western bot 분석법 그리고 면역형광염색법을 수행하였다. 병용처리 된 사람골육종세포는 단독처리 된 사람골육종세포에서 거의 관찰할 수 없었던 핵 응축과 조각남, 사립체막 전위와 DNA 양의 감소, cytochrome c의 세포질로의 유리, AIF의 핵으로의 이동, caspase-3과 caspase-7의 파괴 및 PARP의 분절화와 같은 세포자멸사 증거를 보였다. 48시간 동안 1 mM의 VPA와 0.5 ${\mu}M$ 17AAG을 각기 단독처리 한 결과에서는 세포자멸사를 유도 못했으나, 병용처리한 결과에는 아주 탁월한 세포자멸사의 유도를 보였다. 이러한 병용처리 결과는 사람골육종의 새로운 치료적 전략으로 응용될 수 있다고 생각한다.

• 생명과학회지
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• 제23권11호
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• pp.1351-1359
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• 2013

${\beta}$-Ionone의 인체 비소폐암세포 A-549 cells (human non-small lung cancer A-549 cell)에 대한 anti-proliferative effect에 관한 연구를 수행하였다. A-549 cell에 다양한 농도의 ${\beta}$-ionone (1, 5, 10, and 15 ${\mu}M$)을 2, 4, 6일 간 처리하고, 배양 2일에 A-549 cell의 생육억제와 관련되는 biochemical marker를 측정하였다. ${\beta}$-Ionone은 A-549 cell 생육을 dose와 time 의존적으로 저해하였다. ${\beta}$-Ionone 배양 2일의 $IC_{50}$은 5.0 ${\mu}g/ml$이었다. ${\beta}$-Ionone은 농도 의존적으로 apoptosis를 유도하였다. ${\beta}$-Ionone은 p53, p21 및 Bax protein 수준을 증가시켰으나, Bcl-2 protein 발현은 억제시켰다. ${\beta}$-ionone은 cytosol cytochrome c 함량을 증가시켰고, caspase-9과 caspase-3 효소 활성증가를 유도하였다. 또한, ${\beta}$-ionone은 $cPLA_2$와 COX-2 protein level을 감소시켰다. 이와 같은 결과는 ${\beta}$-ionone의 A-549 cell에 대한 생육억제효과는 Bax와 Bcl-2 gene 발현을 reciprocal regulation하여 유도한 apoptosis와 $cPLA_2$ 및 COX-2 protein 발현 억제에 기인함을 의미한다.

• 동국한의학연구소논문집
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• 제11권
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• pp.52-57
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• 2008

Objectives. In this study, we investigate that methanol extract of Euphorbiae lathyridis Semen contributes to growth inhibitory effect on the HT-29 human colon cancer cells. Methods. Euphorbiae lathyridis Semen (ELS) was extracted with 80% methanol. HT-29 cells were treated with different concentrations of ELS extract for 24-72 hrs. Growth inhibitory effect was determined by MTT assay. Cell apoptosis was determined by surveying caspases cascades activation using Western blot. Cell cycle arrest was analyzed by flow cytometry with PI staining. Results. Exposure to ELS extract showed in inhibitory effects on HT-29 cell growth as a dose-dependent manner. Cell growth inhibition by ELS extract was related with induction of cell apoptosis with DNA fragmentation through the activation of caspases-3, caspase-9 and PARP cleavage. Conclusion. ELS extract significantly inhibited cell growth and induced cell apoptosis in HT-29 human colon cancer cells, therefore, These results suggest that ELS extract can be used as chemoprevention agent of colon cancers.

• Journal of Microbiology and Biotechnology
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• 제28권11호
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• pp.1771-1781
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• 2018

The emergence of multidrug-resistant microorganisms, as well as fungal infectious diseases that further threaten health, especially in immunodeficient populations, is a major global problem. The development of new antifungal agents in clinical trials is inferior to the incidence of drug resistance, and the available antifungal agents are restricted. Their mechanisms aim at certain characteristics of the fungus in order to avoid biological similarities with the host. Synthesis of the cell wall and ergosterol are mainly targeted in clinical use. The need for new approaches to antifungal therapeutic agents or development alternatives has increased. This review explores new perspectives on mechanisms to effectively combat fungal infections and effective antifungal activity. The clinical drug have a common feature that ultimately causes caspase-dependent cell death. The drugs-induced cell death pathway is associated with mitochondrial dysfunction, including mitochondrial membrane depolarization and cytochrome c release. This mechanism of action also reveals antimicrobial peptides, the primary effector molecules of innate systems, to highlight new alternatives. Furthermore, drug combination therapy is suggested as another strategy to combat fungal infection. The proposal for a new approach to antifungal agents is not only important from a basic scientific point of view, but will also assist in the selection of molecules for combination therapy.