• Archives of Pharmacal Research
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• v.25 no.4
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• pp.463-468
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• 2002

We have previously reported on the identification of the endogenous transmethylation inhibitor oligosaccharide-linked acyl carrier protein (O-ACP), In this study, the role of the transmethylation reaction on cell cycle progression was evaluated using various transmethylase inhibitors, including O-ACP. O-ACP significantly inhibited the growth of various cancer cell lines, including NIH3T3, ras-transformed NIH3T3, MDA-MB-231, HT-1376, and AGS. In addition, exposure of ras-transformed NIH3T3 to O-ACP caused cell cycle arrest at the $G_0/G_1$ phase, which led to a decrease in cells at the S phase, as determined by flow cytometry. In contrast, transmethylase inhibitors did not affect the expression of $p21^{WAF1/Cip1}$, a well known inhibitor of cyclin dependent kinase, indicating that the cell cycle arrest by transmethylase inhibitors might be mediated by a $p21^{WAF1/Cip1}$-independent mechanism. Therefore, O-ACP, a novel transmethylase inhibitor, could be a useful tool for elucidating the novel role of methylation in cell proliferation and cell cycle progression.

• BMB Reports
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• v.47 no.7
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• pp.393-398
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• 2014

The role of Bax inhibitor-1 (BI-1) in the protective mechanism against apoptotic stimuli has been studied; however, as little is known about its role in death receptor-mediated cell death, this study was designed to investigate the effect of BI-1 on Fas-induced cell death, and the underlying mechanisms. HT1080 adenocarcinoma cells were cultured in high concentration of glucose media and transfected with vector alone (Neo cells) or BI-1-vector (BI-1 cells), and treated with Fas. In cell viability, apoptosis, and caspase-3 analyses, the BI-1 cells showed enhanced sensitivity to Fas. Fas significantly decreased cytosolic pH in BI-1 cells, compared with Neo cells, and this decrease correlated with BI-1 oligomerization, mitochondrial $Ca^{2+}$ accumulation, and significant inhibition of sodium-hydrogen exchanger (NHE) activity. Compared with Neo cells, a single treatment of BI-1 cells with the NHE inhibitor EIPA or siRNA against NHE significantly increased cell death, which suggests that the viability of BI-1 cells is affected by the maintenance of intracellular pH homeostasis through NHE.

• Journal of Haehwa Medicine
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• v.10 no.2
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• pp.179-188
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• 2002

This experimental study was carried out to evaluate the effects of Acanthopanacis cortex on Cytokine-inducing and and immune response in Mice. In order to investigate the effect of Acanthopanacis cortex, the following was performed; Cytotoxicity, in vitro, the fraction of $CD4^+$, $CD8^+$, $B220^+$ in splenic cell, gene expression of IL-12(p35), IL-12(p40), IFN-${\gamma}$, and splenic cell proliferation by Acanthopanacis cortex. Analysis of cytokine gene expression was carried out by RT-PCR amplification. Amplified PCR products were electrophoresed on 1.2% agarose gel, and the analysis (Ht) was used to 1D-density program. The results were obtained as follows. Acanthpanacis cortex showed didn't have cell toxicity under $12{\mu}g/m{\ell}$ group on mouse lung fibroblast cells. In an in vitro model using mouse peripheral blood mononuclear cells (PBMCs), extract of Acanthpanacis cortex induced multiple cytokine, including interleukin-12 (p35), interleukin-12 (p40), interferon-gamma (IFN-${\gamma}$). The extract also enhanced the percentages of the $CD4^+$, and $CD8^+$ in the untreated control were $22.1{\pm}3.3$ to $38.4{\pm}2.1$, and $5.0{\pm}0.4$ to $10.7{\pm}0.3%$, respectively. From above findings, it is suggested that Acanthopanacis cortex is able to anti-cancer and activate immune response system.

• KSBB Journal
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• v.28 no.5
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• pp.332-337
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• 2013

The gene encoding for xylose isomerase from the thermophilic bacterium Thermotoga maritima was cloned and recombinantly expressed in E. coli cells. Optimal activity was shown at $90^{\circ}C$ and pH 8.0. Treatment of saponin by recombinant xylose isomerase increased the growth inhibitory effect against human gastric cancer (AGS) cells and human colon cancer (HT-29) cells. On the other hand, treatment of fucoidan by the enzyme could not change the growth inhibitory effect against the same cancer cells. One ${\mu}g/ml$ of enzyme-treated saponin exhibited the same or higher growth inhibitory effect against both cancer cells compared with 100 ${\mu}g/ml$ of enzymeuntreated saponin. These results would be useful in the development of functional food or drug.

• Korean Journal of Food Science and Technology
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• v.32 no.6
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• pp.1371-1378
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• 2000

This study was performed to determine the antimutagenic and cytotoxic effect of Agaricus blazei Murill methanol extract on Salmonella typhimurium TA98, TA100 and human cancer cell lines using Ames test and cytotoxicity assay, respectively. In Ames test, methanol extract from A. blazei Murill did not exhibit any mutagenicity and most of the samples showed high antimutagenic effects against mutation induced by N-methyl-N'-nitro-N-nitrosoguanidine(MNNG), 4-nitroquinoline-1-oxide(4NQO), 3-amino-1,4-dimethyl-5H-pyrido [4,3-b] indol(Trp-P-1) and $benzo({\alpha})pyrene(B({\alpha})P)$. The methanol extracts of A. blazei Murill$(200\;{\mu}g/plate)$ showed approximately 92.4%, 81.9% and 83.4% inhibitory effect on the mutagenesis induced by 4NQO, Trp-P-1 and $B({\alpha})P$ against TA98 strain, whereas 87.3%, 94.7%. 92.3% and 89.9% inhibitions were observed on the mutagenesis induced by MNNG, 4NQO, Trp-P-1 and $B({\alpha})P$ against TA100 strain. The solvent fractions of methanol extracts from A. blazei Murill except water fraction showed high antimutagenic effects of $70{\sim}90%$ against mutation induced by MNNG, 4NQO. Trp-P-1 and $B({\alpha})P$. In anticancer effects of A. blazei Murill extract and fraction against cancer cell lines including human breast adenocarcinoma(MCF7), human lung carcinoma(A549), human fibrosarcoma(HT1080), human hepatocellular carcinoma(Hep3B), human epitheloid carcinoma(HeLa), human gastric carcinoma(KATO III) and human chronic myelogenous leukemia(K562) were investigated. The treatment of 1 mg/mL A. blazei Murill extracts had the highest cytotoxicity with 91.9% against HeLa, followed by KATO III(88.7%), A549(86.5%) and Hpe3B(84.3%). Whereas 1 mg/mL treatment of A. blazei Murill extracts had only $10{\sim}40%$ cytotoxicity on human normal liver cell (WRL68).

• BMB Reports
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• v.53 no.9
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• pp.478-483
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• 2020

Kudoa septempunctata is a myxozoan parasite that causes food poisoning in individuals consuming olive flounder. The present study aimed to investigate the currently insufficiently elucidated early molecular mechanisms of inflammatory responses in the intestine owing to parasite ingestion. After Kudoa spores were isolated from olive flounder, HT29 cells were exposed to spores identified to be alive using SYTO-9 and propidium iodide staining or to antigens of Kudoa spores (KsAg). IL-1β, IL-8, TNF-α and NFKB1 expression and NF-κB activation were assessed using real-time PCR, cytokine array and western blotting. The immunofluorescence of FITC-conjugated lectins, results of ligand binding assays using Mincle-Fc and IgG-Fc, CLEC4E expressions in response to KsAg stimulation, and Mincle-dependent NF-κB activation were assessed to clarify the early immune-triggering mechanism. Inflammatory cytokines (IL-1β, GM-CSF and TNF-α), chemokines (IL-8, CCL2, CCL5 and CXCL1) and NF-κB activation (pNF-κB/NF-κB) in HT29 cells increased following stimulation by KsAg. The immunofluorescence results of spores and lectins (concanavalin A and wheat germ agglutinin) suggested the importance of Mincle in molecular recognition between Kudoa spores and intestinal cells. Practically, data for Mincle-Fc and KsAg binding affinity, CLEC4E mRNA expression, Mincle immunofluorescence staining and hMincle-dependent NF-κB activation demonstrated the involvement of Mincle in the early immune-triggering mechanism. The present study newly elucidated that the molecular recognition and immune-triggering mechanism of K. septempunctata are associated with Mincle on human intestinal epithelial cells.

• International Journal of Industrial Entomology and Biomaterials
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• v.8 no.1
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• pp.89-93
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• 2004

Expression of a fusion protein between B. thuringiensis crystal protein, Cry1Ac1 and a scorpion insect toxin (AaIT, Androctonus australis Hector insect toxin) in acrystalliferous B. thuringiensis strain (Cry-B strain) was examined. The cry 1Ac1 gene was cloned in B. thuringiensis-E coli shuttle vector, pHT3101, under the control of the native cry 1Ac1 gene promoter (pProAc) and a gene encoding AaIT was inserted in XhoI site in the middle of the cry 1Ac1 gene (pProAc-ScoR). B. thuringiensis Cry-B strain carrying pProAc-ScoR (PyoAc-ScoR/CB) produced an inclusion body of irregular shape and the expressed fusion protein is approximately 65 kDa in size. Sporulated cells and spore-crystal mixtures of ProAc-ScoR/CB had insecticidal activity against Plutella xylostella larvae, showing $LT_50$ of ProAc-ScoR/CB (22.59 hrs) lower than that of ProAc/CB (30.06 hrs) at $1{\times}{10^7} {CEU/cm^2}$. These results suggest that the fusion protein including a B. thuringiensis crystal protein and an AaIT may be functionally expressed in B. thupingiensis. Moreover, we verified the additive toxicity of AaIT, which is a new feasible candidate for insect control.

• Advances in Traditional Medicine
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• v.6 no.3
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• pp.179-185
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• 2006

We investigated the effects of chronic administration of different extracts of ginger rhizome [pet ether extract (PE); toluene fraction (TF) of pet ether extract] on anxiety models: the elevated T-maze (ETM) (for inhibitory avoidance and escape measurements) and the open field test. Ondansetron (1 mg/kg), FE (10, 30 &100 mg/kg) and TF (10 & 30 mg/kg) were administered orally for 15 days. On the $14^{th}$ day mice were previously exposed for 30 min to one of the open arms of the T-maze, 24 h before the test. On $15^{th}$ day mice had two exposures to the enclosed and open arm of the ETM followed by exposure to the open field apparatus. The number of line crossings in the apparatus was used to assess locomotor changes. Cumulative Concentration Response Curve of 5-HT was plotted using rat fundus which were pretreated in a similar way. Treatment with Ondansetron (1 mg/kg), PE (100 mg/kg), TF (10 mg/kg) and TF (30 mg/kg) significantly (P<0.05) impaired inhibitory avoidance performance but did not impair escape latency. Concentration response curve of 5-HT was shifted towards the right with suppression of maxima in rats treated with PE and TF. The results suggest that PE and TF of Ginger rhizome exerts anxiolytic like behaviour in a specific subset of defensive behaviour, particularly those related to generalized anxiety disorder.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.12
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• pp.1915-1923
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• 2013

The aim of this study is carried out to evaluate the fermentation characteristics and increased functionality when doenjang was prepared with bamboo salts. Grain type mejus were fermented with mixed starter cultures of Aspergillus oryzae, Bacillus subtilis, and Lactococcus lactis. These mejus were mixed with different kinds of salts-purified salt (PD), sea salt (SD), one-time baked bamboo salt ($1{\times}BD$), three-time baked bamboo salt ($3{\times}BD$), and nine-time baked bamboo salt ($9{\times}BD$)- when doenjangs were prepared. For doenjang fermentation period of 8 weeks at $37^{\circ}C$, the fermentation characteristics of all the groups were compared. The amino type nitrogen content and enzyme activities (protease and ${\alpha}$-amylase) in the samples were significantly increased. In DPPH radical scavenging activities and hydroxyl radical scavenging activities, $9{\times}BD$ (47% and 69%) showed the highest scavenging activities compared to PD (40% and 49%), SD (42% and 57%), $1{\times}BD$ (42% and 64%) and $3{\times}BD$ (45% and 65%) (P<0.05). The anticancer effects of doenjang in HT-29 cancer cells indicated all the groups, especially doenjang prepared with bamboo salts were higher than the others (P<0.05). Apoptosis related genes of Bax and Bcl-2, as well as inflammation related genes of iNOS and COX-2 were regulated by the treatment of doenjangs in HT-29 cancer cells. SD, $1{\times}BD$, $3{\times}BD$, and $9{\times}BD$ increased the expression level of Bax and decreased the expression level of Bcl-2, iNOS, and COX-2. These results suggest that sea salt and bamboo salt especially bamboo salt could improve fermentation characteristics and functionality of doenjang and play an important role in regulating apoptosis and inflammation related genes in cancer cells.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.1
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• pp.299-302
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• 2013

Methylenetetrahydrofolate reductase (MTHFR) has been reported to be associated with DNA methylation, an epigenetic feature frequently found in gastric cancer. We conducted a case-control study to explore the association of MTHFR C677T polymorphisms with gastric cancer risk and its relation with the DNA methylation of COX-2, MGMT, and hMLH1 genes. Genotyping of P16, MGMT and HMLH1 was determined by methylation-specific PCR after sodium bisulfate modification of DNA, and genotyping of MTHFR C677T was conducted by TaqMan assays using the ABI Prism 7911HT Sequence Detection System. Folate intake was calculated with the aid of a questionnaire. Compared with the MTHFR 677CC genotype, the TT genotype was significantly associated with 2.08 fold risk of gastric cancer when adjusting for potential risk factors. Individuals who had an intake of folate above $310{\mu}g$/day showed protective effects against gastric cancer risk. The effect of MTHFR C677T polymorphisms on the risk of gastric cancer was modified by folate intake and methylation status of MGMT (P for interaction <0.05).