• Journal of Life Science
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• v.27 no.3
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• pp.267-274
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• 2017

The patient with end-stage renal disease show several nervous complications. The factors contributing to the nervous complications are still incompletely characterized. Cyanate, known as one of the uremic toxins, is derived spontaneously from urea. To investigate the mechanism of cyanate-induced effect on C6 glioma cells, the glioma cells were treated with 0, 1, 5, 10, 20 and 40 mM cyanate. There was a dose-dependent decrease in cell viability and the decreased number of cell was observed in glioma cells by treatment with cyanate. Western blot showed the down- regulation of procaspase-3, which means up-regulation of caspase-3, and the up-regulation of caspase-8, but the down-regulation by cyanate. In addition, cDNA microarray showed 934 down-regulated genes and 165 up-regulated genes on 1,099 genes in cyanate treated group. Treatment with cyanate led to 16 down-regulated genes and 6 up-regulated genes on apoptosis category, and especially heat shock 70 kD protein 1A gene on the category of apoptosis was significantly up-regulated. These results suggest that cyanate can induce apoptosis through caspase-8 and caspase-3 in glioma cells and decrease of gene expression including apoptosis category in glioma cells. These effects of cyanate may play a role in the nervous complications of patient with end-stage renal disease.

• Nutrition Research and Practice
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• v.11 no.2
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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.

• Korean Journal of Microbiology
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• v.42 no.4
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• pp.319-325
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• 2006

[ $\beta$ ]-Glucans (AG) were prepared from Agaricus blazei cultured in the medium fortified with the roots of Pueraria spp. by repeated extraction with hot water, gel filtration chromatography and DEAE ion exchange chromatography. Oligosaccharides (AO) were derived from the hydrolysis of AG by an endo-$\beta$-(1$\rightarrow$6)-glucanase from Bacillus megaterium. The anti-HT-29 human colon cancer activity of AG or AO was investigated using MTT assay, apoptosis assay, cell cycle analysis, and cDNA microairay. AG and AO both inhibited proliferation and growth of HT-29 cells, and stimulated apoptosis of the cells in a dose-dependent manner. In cell cycle analysis, treating HT-29 cells with AG or AO resulted in the increase of cells in the G0 (sub-G1) and G1 phase. Especially, AO was more effective in inducing G0/G1 cell cycle arrest than AG. To screen the genes involved in the increase of apoptosis, the gene expression profile of the HT-29 cells treated with AO was examined by cDNA microarray. While several genes involved in cell cycle progression (CCND2 and CDK2) were down-regulated, many genes involved in apoptosis (TNFSF9, TNFRSF9, FADD, CASP8, BAD, CRADD, CASP9 etc), cell cycle inhibitor (CDKN2A), immune response (IL6, IL18, IL6R etc), and tumor suppressor (CEACAM1, TP53BP2, IRF1, and PHB) were up-regulated. These results suggest that AO could inhibit the proliferation and growth of HT-29 cells by G0/G1 cell cycle arrest and induction of apoptosis.

• Journal of Life Science
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• v.22 no.12
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• pp.1672-1679
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• 2012

The effect of high stocking density (HSD) on the expression of stress and lipid metabolism associated genes in the liver of broiler chickens was examined by chicken genome array analysis. The chickens in a control group were randomly assigned to a $495cm^2/bird$ stocking density, whereas the chickens in a HSD group were arranged in a $245cm^2/bird$ stocking density with feeding ad libitum for 35 days. The chickens assigned to the HSD group had a significantly lower body weight, weight gain, and feed intake compared with those of the control group (p<0.05). The mortality of chickens was higher in the HSD group than in the control group. The microarray analysis indicated up-regulation of stress associated genes such as HMGCR, $HSP90{\alpha}$, HSPA5 (GRP78/Bip), DNAJC3 and ATF4, and down-regulation of interferon-${\gamma}$ and PDCD4 genes. The endoplasmic reticulum stress associated genes, HSPA5 (GRP78/Bip), DNAJC3 and ATF4, were highly expressed in the HSD group. The genes, ACSL5, TMEM195 and ELOVL6, involved in fatty acid synthesis, were elevated in the HSD group. The genes, ACAA1, ACOX1, EHHADH, LOC423347 and CPT1A, related to fatty acid oxidation, were also activated in the HSD group. These results suggest that a HSD rearing system stimulates the genes associated with fatty acid synthesis as well as fatty acid oxidation in the liver of broiler chickens.

• Microbiology and Biotechnology Letters
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• v.40 no.2
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• pp.117-127
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• 2012

The beneficial effects of adipose-derived stem cell conditioned media (ADSC-CM) for skin regeneration have previously been reported, despite the precise mechanism of how ADSC-CM promotes skin regeneration remaining unclear. ADSC-CM contains various secretomes and this may be a factor in it being a good resource for the treatment of skin conditions. It is also known that ADSC-CM produced in hypoxia conditions, in other words Advanced Adipose-Derived Stem cell Protein Extract (AAPE), has excellent skin regenerative properties. In this study, a human primary skin cell was devised to examine how AAPE affects human dermal fibroblast (HDF) and human keratinocyte (HK), which both play fundamental roles in skin regeneration. The promotion of collagen formation by HDFs was observed at 0.32 mg/ml of AAPE. AAPE treatment significantly stimulated stress fiber formation. DNA gene chips demonstrated that AAPE in HKs (p<0.05) affected the expression of 133 identifiable transcripts, which were associated with cell proliferation, migration, cell adhesion, and response to wounding. Twenty five identified proteins, including MMP, growth factor and cytokines such as CD54, FGF-2, GM-CSF, IL-4, IL-6, VEGF, TGF-${\beta}2$, TGF-${\beta}3$, MMP-1, MMP-10, and MMP-19, were contained in AAPE via antibody arrays. Thus, AAPE might activate the HK biological function and induce the collagen synthesis of HDF. These results demonstrate that AAPE has the potential to be used for clinic applications aimed at skin regeneration.

• Journal of Life Science
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• v.22 no.1
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• pp.49-54
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• 2012

The effects of genistein on cell proliferation and adipogenesis were examined in mouse 3T3-L1 preadipocyte cells. Genistein decreased viability of 3T3-L1 pre-adipocytes in a dose-dependent manner. Oil Red O staining of these cells also indicated that adipogenesis was inhibited by 50 ${\mu}M$ genistein treatment. We investigated the molecular mechanisms involved in the decrease in cell viability in genistein-treated 3T3-L1 cells by conducting an oligo DNA microarray analysis. We selected the sirtuin-1 gene, one of the upregulated genes, for further experimentation because sirtuin-1 belongs to the sirtuin family, which is associated with anti-obesity and anti-inflammation activities. We found that four phytochemicals (resveratrol, capsaicin, daidzein, and genistein) could increase sirtuin-1 expression. Genistein was the strongest inducer of sirtuin-1 among the tested phytochemicals. The inhibition of adipogenesis by genistein was recovered by surtuin-1 siRNA transfection. Overall, these results may further our understanding of the molecular mechanisms underlying the inhibition of proliferation and adipogenesis by genistein in mouse 3T3-L1 cells.

• Proceedings of the Korean Society for Bioinformatics Conference
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• 2006.02a
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• pp.83-89
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• 2006

Cellular functions are carried out by a concerted action of biochemical pathways whose components have genetic interactions. Abnormalities in the activity of the genes that constitute or modulate these pathways frequently have oncogenic implications. Therefore, identifying the upstream regulatory genes for major biochemical pathways and defining their roles in carcinogenesis can have important consequences in establishing an effective target-oriented antitumor strategy We have analyzed the gene expression profiles of human liver cancer samples using cDNA microarray chips enriched in liver and/or stomach-expressed cDNA elements, and identified groups of genes that can tell tumors from non-tumors or normal liver, or classify tumors according to clinical parameters such as tumor grade, age, and inflammation grade. We also set up a high-throughput cell-based assay system (cell chip) that can monitor the activity of major biochemical pathways through a reporter assay. Then, we applied the cell chip platform for the analysis of the HCC-associated genes discovered from transcriptome profiling, and found a number of cancer marker genes having a potential of modulating the activity of cancer-related biochemical pathways such as E2F, TCF, p53, Stat, Smad, AP-1, c-Myc, HIF and NF-kB. Some of these marker genes were previously blown to modulate these pathways, while most of the others not. Upon a fast-track phenotype analysis, a subset of the genes showed increased colony forming abilities in soft agar and altered cell morphology or adherence characteristics in the presence of purified matrix proteins. We are currently analyzing these selected marker genes in more detail for their effects on various biological Processes and for Possible clinical roles in liver cancer development.

• Journal of Acupuncture Research
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• v.25 no.2
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• pp.11-26
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• 2008

목적 : 홍삼약침(藥鍼)이 고혈당 및 지질대사장애에 미치는 개선효과와 그 기전을 조사하고자 한다. 방법 : 홍삼약침(藥鍼)의 anti-diabetic 활성과 그 기전을 C57BL/KsJ db/db mice를 이용하여 관찰하였다. 실험 동물은 대조군(DC), 홍삼약침(藥鍼)군(RGL, RGH) 및 양성대조군(MET, GPZ, PIO)의 6군으로 나누었다. 홍삼약침(藥鍼)군은 $0.2m{\ell}$의 홍삼약침멸(藥鍼滅)을 각각 100mg/kg(RGL) 및 200mg/kg(RGH)씩 인체의 간유(肝兪)($BL_{18}$)에 상응하는 혈위에 1일 1회 10주간 좌우 혈을 번갈아가며 약침 시술하였다. 양성대조군은 metformin 300mg/kg(MET), glipizide 15mg/kg(GPZ) 및 pioglitazone 30mg/kg(PIO)을 각각 1일 1회 10주간 경구투여 하였다. 체중과 혈당은 매주 측정하였다. 실험 10주 후에는 혈액채취로 혈중 glucose, 당화혈색소(HbAlc), insulin, 중성지방(TG), adiponectin, leptin, non-esterified fatty acid(NEFA)를 측정 하였고, 간 조직을 채취하여 조직학적 검사 및 gene expression 분석을 시행하였다. 결과 : 홍삼약침(藥鍼)(RGL, RGH)은 10주 동안 C57BL/KsJ db/db mice의 체중을 증가시키는 부작용은 나타나지 않았다. 홍삼약침(藥鍼)군(RGL, RGH)의 사료섭취량은 대조군과 비슷하였으나 음수량은 증가하였다. 홍삼약침(藥鍼)(RGL, RGH)은 대조군에 비하여 각각 19.8% 및 18.3% 혈당을 낮추었고, 홍삼약침(藥鍼)(RGL)은 insulin resistance를 27.7% 감소시켰으며, 경구내당능 검사의 혈중 glucose에서는 대조군에 비해 홍삼약침(藥鍼)군(RGL, RGH)과 양성대조군(MET, GPZ, PIO)에서 각각 19.8%, 18.3%, 67.7%, 52.3% 및 56.9% 감소시켰다. 당화혈색소(HbAlc)는 홍삼약침(藥鍼)(RGL, RGH), MET, GPZ 및 PIO군에서 대조군에 비하여 각각 11.0%, 6.4%, 18.9%, 16.1% 및 27.9% 감소시켰으며, 혈중 glucose감소와 유사한 경향을 나타내었다. 홍삼약침(藥鍼)(RGL)은 대조군에 비해 TG와 NEFA를 각각 18.8% 및 16.8% 감소시켰고, adiponectin과 leptin을 각각 20.6% 및 12.1% 증가시켰다. 홍삼약침(藥鍼)(RGL, RGH)은 중성지방의 침착으로 인한 간의 질량비 증가를 억제하지 못하였으나, 지방구를 감소시겼음을 관찰할 수 있었다. Microarray 분석에서는 홍삼약침(藥鍼)(RGL, RGH)이 간에서 glycolysis, gluconeogenesis 및 fatty acid beta-oxidation과 관련된 유전자 발현에 영향을 미치는 것으로 나타나 양성대조군 metformin과 유사한 기전을 나타내었다. 요약 : 홍삼약침(藥鍼)은 T2DM동물모델(C57BL/KsJ db/db mice)에서 항당뇨 및 지질대사 개선활성이 있었다. 홍삼약침(藥鍼)은 C57BL/KsJ db/db mice의 간조직에서 lipogenesis억제 및 fatty acid beta-oxidation활성을 통해 혈당 이용을 높이고, insulin sensitivity를 향상시켰다. 또한 유전자 발현분석을 통해 그 기전이 metformin과 유사함을 확인할 수 있었으므로 향후 홍삼약침(藥鍼)의 새로운 약침 기술 개발 근거가 될 수 있을 것으로 사료된다.

• Korean Journal of Acupuncture
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• v.24 no.2
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• pp.129-139
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• 2007

목적 : Signal transducers and activators of transcription 6 (Stat 6) 유전자는 면역세포의 발달에 있어서 중요한 유전인자이며, IL-4와 같은 사이토카인에 의해 유전자 발현이 조절된다. 본 연구에서는 Stat 6 유전자 제거 생쥐와 정상 (wild type, W/T) 생쥐에 carrageenan으로 염증을 유도한 후 족삼리에 침치료를 시행하여 시상하부에서의 유전자 발현 양상을 분석하고자 하였다. 방 법 : BALB/c (W/T, n=12) and BALB/c-Stat 6 유전자 제거 생쥐 (n=12)의 발뒤꿈치 표피에 1% carrageenan을 30 ul 주사하여 염증을 유도하였다. 침은 염증 유도 30분 후에 족삼리(ST36)에 시침하였으며, 염증유도에 의한 부종 증가율을 매 시간마다 측정하여 총 5시간동안 측정하였다. 마이크로에러이는 Stat 6 유전자 제거 생쥐를 염증 유발 군과 염증유발 후 침을 처치한 군으로 나누고, 시상하부를 적출하여 RNA를 분리한뒤 마이크로어레이 프로파일을 분석하였다. 결 과 : 염증에 의한 부종증가율을 비교한 결과, Stat 6 유전자 제거 생쥐 그룹의 부종증가율이 W/T 생쥐의 부종 증가율보다 약 50 % 정도 감소하였으며, 각 3, 4, 5시간째에 유의한 차이를 나타내었다. (각 p<0.05). W.T생쥐군과 Stat 6 유전자 제거 생쥐군 모두에서, 침 처치군이 염증 유발 군에 비해, 염증 유발 2시간 후부터 유의한 감소를 나타내었다. 시상하부의 유전자 발현을 관찰한 결과, 39개의 유전자가 3배 이상 감소하였으며, 19개의 유전자는 3배 이상 증가하였다. 결 론 : W/T 생쥐군과 Stat 6 유전자 제거 생쥐 모두에서 침의 진통효과는 나타나며, 이의 기전에는 시상하부에서의 침 치료에 의한 염증관련 유전자들의 감소와, 항염증과 관련된 유전자들이 증가가 관여하는 것으로 보인다., 10, 11), 내측전완피신경(TE5, 6, 7, 8, 9, 10, 11), 후상완피신경(TE12, 13), 상외측상완피신경(TE13), 외측쇄골상신경(TE14, 15),대이개신경(TE16, 17, 18, 19), 소후두신경(TE19, 20), 이개측두신경(TE20, 21, 22), 안면신경측두지(TE22, 23), 관골측두신경(TE23), 중층에 견갑상신경(TE15), 견갑배신경(TE15), 경상설골근신경(TE17), 후이개신경(TE18, 19, 20), 안면신경측두지(TE20, 21, 22), 심층에 후골간신경(TE5, 6, 7), 요골신경심지(TE8, 9, 12, 13), 견갑상신경(TE14), 액와신경가지(TE14), 부신경(TE16), 안면신경과 부신경가지(TE17), 설인신경(TE17), 설하신경(TE17), 경신경고리(TE17), 미주신경(TE17), 안면신경 (TE18). 3) 혈(血) 관(管) : 천층에 척측정맥배측지(TE1, 2), 고유수장지동맥배측지(TE1), 배측중수골동맥배측지(TE2), 배측중수골정맥(TE3), 척측피정맥(TE4, 5, 6, 7, 8, 9, 10, 11), 배측정맥궁(TE4), 부요측피정맥(TE6, 8, 9),요측피정맥(TE10, 11), 후견봉정맥가지(TE13, 14), 후이개동 ${\cdot}$ 정맥(TE16, 17, 18, 19, 20), 전이개동 ${\cdot}$ 정맥(TE20), 천측두동 ${\cdot}$ 정맥(TE22, 23), 중층에 후상완회선동맥(TE14), 견갑배동맥(TE15), 견갑상동맥(TE15),천측두동 ${\cdot}$ 정맥(TE21), 관골측두동 ${\cdot}$ 정맥(TE23), 심층에 배측중수골동맥(TE3), 배측수근동맥궁(TE4), 후골간동맥(TE4, 5, 6, 7, 8, 9), 전골간동맥(TE6, 7, 9)

• Maxillofacial Plastic and Reconstructive Surgery
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• v.28 no.2
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• pp.95-110
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• 2006

The treatment for oral and maxillofacial carcinoma with chemotherapeutic agents is evaluated by many effective methods to reduce the tumor mass and cancer cell proliferation. However these chemotherapy have many serious side effects, such as bone marrow suppression, renal toxicity, G-I troubles. Therefore a possible approach to develop a clinically applicable chemotherapeutic agent is to screen anticancer activity of Taxol which is known to have very little side effect and have been used to breast cancer and ovarian carcinoma. Taxol is a new anti-microtubular anti-cancer agent extracted from the bark of the Pacific yew, Taxus brevifolia. Paclitaxel(Taxol) acts by promoting tubulin polymerization and over stabilizing microtubules agianst depolymerization. Despite the constant improvements of methods of the cancer treatment especially chemotherapy, the rate of cancer metastasis and recurrent are not decreased. Thus the investigation of new drug which have very little side effect and a possible clinically application continues to be a high priority. Considering that the Taxol have shown very effective chemotherapeutic agent with relatively low toxicity in many solid tumors, it deserves to evaluate its efficacy in oral squamous cell carcinoma. In this study, to investigate the in-vivo and in-vitro anti-cancer efficacy of Taxol in oral squamous cell carcinoma and lastly, the potency of Paclitaxel in the clinical application for oral cancer was evaluated. In vivo study, after HN22 cell line were xenografted in nude mice, the growth of tumor mass was observed, 3 mg/Kg taxol was injected intraperitoneally into nude mice containing tumor mass. The methods of these study were measurement of total volume of tumor mass, histopathologic study, immunohistochemical study, drug resistance assay, growth curve, MTT assay, flow cytometry, cDNA microarray in vivo and in vitro. The results were obtained as following. 1. The visual inspection of the experimental group showed that the volume of the tumor mass was slightly decreased but no significant difference with control group. 2. Ki-67 index was decreased at weeks 4 in experimental group. 3. Microscopic view of the xenografted tumor mass showed well differentiated squamous cell carcinoma and after Taxol injection, some necrotic tissue was seen weeks 4. 4. The growth curve of the tumor cells were decreased after 1day Taxol treatment. 5. According to the MTT assay, HN22 cell line showed relative drug resistancy above $5\;{\mu}g/ml$ concentrations of Taxol. 6. In drug resistance assay, the decrease of cell counts was seen relatively according to concentration. 7. In Flow cytometry, G2M phase cell arrests were seen in low concentration of the Taxol, while S phase cell arrests were seen in high concentration of the Taxol. 8. Using cDNA microarray technique, variable gene expression of ANGPTL4, TXNRD1, FAS, RRAGA, CTGF, CYCLINEA, P19, DUSP5, CEBPG, BTG1 were detacted in the oral squamous cell carcinoma cell after taxol treatment. In this study paclitaxel is effective against oral squamous cell carcinoma cell lines in vitro, but week effect was observed in vivo. So we need continuous study about anticancer effect of taxol in vivo in oral squamous cell carcinoma.