• Title/Summary/Keyword: EBP2

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Evaluation of the Immunohistochemical Staining Pattern of the mTOR Signaling Proteins in Colorectal Cancers and Adenoma Lesions (대장암과 선종 병변에서 mTOR 신호 단백질의 면역조직화학 염색성 평가)

  • Kim, Jin Mok;Lee, Hyoun Wook
    • Korean Journal of Clinical Laboratory Science
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    • v.49 no.4
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    • pp.470-476
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    • 2017
  • Changes in the mammalian target of the rapamycin (mTOR) signaling proteins have been observed in many types of cancer. Accordingly, these proteins have recently become an exciting new target for molecular therapeutics. This study examined the expression of an activated mTOR signaling protein in patients with colorectal adenocarcinoma (CRAC) and colorectal adenoma lesion. Immunohistochemical analysis was performed on human CRAC and adenoma for the mTOR signaling components, including mTOR, phosphorylation, and activation of S6 kinase (p70-S6K), S6 ribosomal protein (S6), and eukaryotic initiation factor 4E-binding protein (4EBP1). A total of 100 cases with colorectal adenocarcinoma (CARC; N=40), adenoma with high-grade intraepithelial neoplasms (HIN; N=30), and adenoma with low-grade intraepithelial neoplasms (LIN; N=30) were enrolled in this study. p-mTOR expression was observed in 30 cases of the CRAC tissues (75%), 9 cases of adenoma with HIN (30%), and 2 cases of adenoma with LIN (7%). In addition, p-S6 expression was observed in 22 cases of CRAC tissues (55%), 8 cases of adenoma with HIN (27%), and 3 cases of adenoma with LIN (10%). A significant correlation was observed among the p-mTOR, p-S6 expression, and the adenoma-carcinoma sequence. Interestingly, the p-S6 protein was activated more in early CRAC than in advanced CRAC.

Effect of hot water and ethanol extracts from Nelumbo nucifera Gaertner flower on lipid accumulation and reactive oxygen species (ROS) production in adipogenesis of 3T3-L1 cells (3T3-L1 세포분화 중 지방축적 및 ROS 생성에 대한 연화 열수 및 에탄올 추출물의 효과)

  • Oh, Ji-Won;Lee, Jin-Ha;Lee, Ok-Hwan;Kim, Kye-Hoon;Kim, Hey-Ran;Lee, Hyo-Ku
    • Food Science and Preservation
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    • v.22 no.5
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    • pp.744-750
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    • 2015
  • The present study was designed to investigate the effects of hot water and ethanol extracts of Nelumbo nucifera Gaertner flower on lipid accumulation and reactive oxygen species (ROS) production during adipogenesis in 3T3-L1 cells. 3T3-L1 preadipocytes were treated with both hot water and ethanol extracts for up to 8 days following standard induction of differentiation. Regarding anti-adipogenic activity, compared with the control, the hot water and ethanol extracts significantly inhibited lipid accumulation (37.4 and 66.6%, respectively) and ROS production (46.4 and 46.8%, respectively) during adipogenesis in 3T3-L1 cells. Treatment with hot water and ethanol extracts significantly inhibited mRNA expression of peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) and CCAAT/enhancer-binding protein alpha ($C/EBP{\alpha}$), thereby reducing the mRNA expression of adipocyte-specific fatty acid binding protein (aP2). Moreover, both the extracts significantly inhibited mRNA expression of NADPH oxidase (NOX4). Overall, our research suggests that N. nucifera Gaertner flower extracts might be a valuable source of bioactive compounds that exhibit anti-adipogenic activity and could have applications in the field of medicine and food industry.

Low Lysine Treatment Increases Adipogenic Potential of Bovine Intramuscular Preadipocytes

  • Beloor, Jagadish;Kang, Hye Kyeong;Yun, Cheol-Heui;Kim, Sang Hoon;Moon, Yang Soo
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.5
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    • pp.721-726
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    • 2009
  • The molecular mechanism of adipocyte differentiation has been well documented. However, the effect of specific nutrients such as lysine on adipocyte differentiation is poorly understood especially in ruminant animals. Therefore, the aim of the present study was to elucidate the influence of lysine on adipocyte differentiation and adipogenic genes in cultured bovine preadipocytes. The preadipocytes were treated with different concentrations of lysine (40, 160, 320 mg/L) or troglitazone (10 ${\mu}M$) for 2 days and then subsequently cultured in differentiation medium until day 6. Expression levels of $C/EBP{\alpha}$ were significantly higher (p<0.001) in 40 and 160 mg/L lysine-treated cells compared to 320 mg/L treatment. Though there was an increasing trend in $PPAR{\gamma}$ expression levels with the decreasing lysine concentration, the results were not significant. The preadipocyte factor (pref-1), expression significantly (p<0.001) reduced with decreasing lysine concentration. The Oil red O staining results were better in 40 mg/L treated cells compared to 160 and 320 mg/L lysine treated cells. Our overall results indicate that insufficient supply of lysine increases the adipogenic potential in bovine intramuscular preadipocytes.

Simvastatin Induces Osteogenic Differentiation and Suppresses Adipogenic Differentiation in Primarily Cultured Human Adipose-Derived Stem Cells

  • Sun, So-Hyun;Lee, Il-Kyu;Lee, Jee-Won;Shim, In-Sop;Kim, Se-Hong;Kim, Kyung-Soo
    • Biomolecules & Therapeutics
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    • v.17 no.4
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    • pp.353-361
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    • 2009
  • Recent in vitro and in vivo animal studies have reported that statin, a cholesterol-lowering drug, stimulate osteogenic differentiation. In the present study, we investigated the effect of simvastatin on osteogenic and adipogenic differentiation in primarily cultured human adipose-derived stem cells (hADSCs). The simvastatin treatment significantly increased the positive cell numbers in alkaline phosphatase and von Kossa staining, and enhanced the expression levels of bone morphogenic protein (BMP)-2, core binding factor alpha 1 (cbfa1), collgen type I and osteonectin mRNAs. Lastly, hADSCs were cultured in the adipogenic media with or without simvastatin to examine the effect of simvastatin on adipogenic differentiation. In the RT-PCR analysis, there were notable decreases in mRNA expression of aP1, C/EBP-$\alpha$ and PPAR-$\gamma$ in hADSCs cultivated in simvastatin-added medium, compared to those in simvastatin-free medium. It suggests that the adipogenic differentiation was significantly inhibited by simvastatin treatment. These observations indicate that simvastatin induces osteogenic differentiation and suppresses adipogenic differentiation in hADSCs.

Emergy Cost-Benefit Evaluation of the Down Stream of Nakdong River Using Environmental-Ecological Concept (환경 생태학적 개념을 이용한 낙동강 하류의 에머지 비용-편익 평가)

  • Jung, Hwa-Sook;Lee, Seog-Mo;Son, Hyeng-Sik;Son, Hee-Jong
    • Journal of Environmental Science International
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    • v.22 no.4
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    • pp.507-514
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    • 2013
  • The Nakdong River being used as drinking water sources for the Busan metropolitan city has the vulnerability of water management due to the fact that industrial areas are located in the upper Nakdong River. This study used emergy analysis method to evaluate ecological-economics of water treatment systems of D water treatment plant (WTP) where located in the downstream of the Nakdong River. The emergy methodology is a system evaluation tool that uses energy as the common currency to compare different resources on a common basis. Emergy yield ratio (EYR) and emergy sustainability index (EmSI) of D WTP were 1.16 and 0.18, respectively. It means not resources and sustainable system but consumer goods and not sustainable system. Ratio of emergy benefit to the purchaser (EBP) shows 2.7 times higher than economic costs. To change the weak water source and situations we need to diversity water intake.

Dlx3 and Dlx5 Inhibit Adipogenic Differentiation of Human Dental Pulp Stem Cells

  • Lee, Hye-Lim;Nam, Hyun;Lee, Gene;Baek, Jeong-Hwa
    • International Journal of Oral Biology
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    • v.37 no.1
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    • pp.31-36
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    • 2012
  • Dlx3 and Dlx5 are homeobox domain proteins and are well-known regulators of osteoblastic differentiation. Since possible reciprocal relationships between osteogenic and adipogenic differentiation in mesenchymal stem cells exist, we examined the regulatory role of Dlx3 and Dlx5 on adipogenic differentiation using human dental pulp stem cells. Over-expression of Dlx3 and Dlx5 stimulated osteogenic differentiation but inhibited adipogenic differentiation of human dental pulp stem cells. Dlx3 and Dlx5 suppressed the expression of adipogenic marker genes such as $C/EBP{\alpha}$, $PPAR{\gamma}$, aP2 and lipoprotein lipase. Adipogenic stimuli suppressed the mRNA levels of Dlx3 and Dlx5, whereas osteogenic stimuli enhanced the expression of Dlx3 and Dlx5 in 3T3-L1 preadipocytes. These results suggest that Dlx3 and Dlx5 exert a stimulatory effect on osteogenic differentiation of stem cells through the inhibition of adipogenic differentiation as well as direct stimulation.

Epigenetic Silencing of CHOP Expression by the Histone Methyltransferase EHMT1 Regulates Apoptosis in Colorectal Cancer Cells

  • Kim, Kwangho;Ryu, Tae Young;Lee, Jinkwon;Son, Mi-Young;Kim, Dae-Soo;Kim, Sang Kyum;Cho, Hyun-Soo
    • Molecules and Cells
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    • v.45 no.9
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    • pp.622-630
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    • 2022
  • Colorectal cancer (CRC) has a high mortality rate among cancers worldwide. To reduce this mortality rate, chemotherapy (5-fluorouracil, oxaliplatin, and irinotecan) or targeted therapy (bevacizumab, cetuximab, and panitumumab) has been used to treat CRC. However, due to various side effects and poor responses to CRC treatment, novel therapeutic targets for drug development are needed. In this study, we identified the overexpression of EHMT1 in CRC using RNA sequencing (RNA-seq) data derived from TCGA, and we observed that knocking down EHMT1 expression suppressed cell growth by inducing cell apoptosis in CRC cell lines. In Gene Ontology (GO) term analysis using RNA-seq data, apoptosis-related terms were enriched after EHMT1 knockdown. Moreover, we identified the CHOP gene as a direct target of EHMT1 using a ChIP (chromatin immunoprecipitation) assay with an anti-histone 3 lysine 9 dimethylation (H3K9me2) antibody. Finally, after cotransfection with siEHMT1 and siCHOP, we again confirmed that CHOP-mediated cell apoptosis was induced by EHMT1 knockdown. Our findings reveal that EHMT1 plays a key role in regulating CRC cell apoptosis, suggesting that EHMT1 may be a therapeutic target for the development of cancer inhibitors.

2, 4-Thiazolidindion Induced Plasticity of Myoblast (C2C12) and Satellite Cells (Porcine) - A Comparative Study

  • Singh, N.K.;Chae, H.S.;Hwang, I.H.;Yoo, Y.M.;Ahn, C.N.;Lee, H.J.;Park, H.J.;Chung, H.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.7
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    • pp.1115-1119
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    • 2007
  • This study was conducted to determine the difference between satellite cells (porcine) and myoblasts (C2C12) in their differentiation under the influence of 2, 4-thiazolidindion. C2C12 myoblast cells and porcine satellite cells (isolated from 10 d old $Landrace{\times}Duroc$ piglets) were grown to absolute confluency. Post confluent cells (day 0) were further exposed to adipogenic induction medium along with 2, 4-thiazolidindion ($8{\mu}M$) for 2 d. Thereafter, cells were exposed to 2, 4-thiazolidindion alone every 2 d till day 10 and analysed. The control was cultured in differentiation medium without any treatment. Increased (p<0.05) expression of transcriptional factors i.e. C/EBP-${\alpha}$ and PPAR-${\gamma}$ and transition of cells to adipocyte morphology was noticed from 2 d and 4 d onwards in satellite cells (Porcine) and myoblasts (C2C12) respectively. Myogenesis was observed to be suppressed completely in case of satellite cells compared to myoblasts in response to 2, 4-thiazolidindion. Pax-7 (transcriptional factor) appeared as a sole entity to satellite cells only, as it was not identified in case of myoblasts. Although both the cells were converting to adipoblasts, the degree of their conversion was different in response to 2, 4-thiazolidindion. Therefore, the hypothesis that satellite cells contribute various domains to the growing myoblasts appeared obscured and found to be dependent on the proliferative energy/or degree of fusion. However, it revealed satellite cells as currency to myoblasts/muscle.

Characterization of H460R, a Radioresistant Human Lung Cancer Cell Line, and Involvement of Syntrophin Beta 2 (SNTB2) in Radioresistance

  • Im, Chang-Nim;Kim, Byeong Mo;Moon, Eun-Yi;Hong, Da-Won;Park, Joung Whan;Hong, Sung Hee
    • Genomics & Informatics
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    • v.11 no.4
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    • pp.245-253
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    • 2013
  • A radioresistant cell line was established by fractionated ionizing radiation (IR) and assessed by a clonogenic assay, flow cytometry, and Western blot analysis, as well as zymography and a wound healing assay. Microarray was performed to profile global expression and to search for differentially expressed genes (DEGs) in response to IR. H460R cells demonstrated increased cell scattering and acidic vesicular organelles compared with parental cells. Concomitantly, H460R cells showed characteristics of increased migration and matrix metalloproteinase activity. In addition, H460R cells were resistant to IR, exhibiting reduced expression levels of ionizing responsive proteins (p-p53 and ${\gamma}$-H2AX); apoptosis-related molecules, such as cleaved poly(ADP ribose) polymerase; and endoplasmic reticulum stress-related molecules, such as glucose-regulated protein (GRP78) and C/EBP-homologous protein compared with parental cells, whereas the expression of anti-apoptotic X-linked inhibitor of apoptosis protein was increased. Among DEGs, syntrophin beta 2 (SNTB2) significantly increased in H460R cells in response to IR. Knockdown of SNTB2 by siRNA was more sensitive than the control after IR exposure in H460, H460R, and H1299 cells. Our study suggests that H460R cells have differential properties, including cell morphology, potential for metastasis, and resistance to IR, compared with parental cells. In addition, SNTB2 may play an important role in radioresistance. H460R cells could be helpful in in vitro systems for elucidating the molecular mechanisms of and discovering drugs to overcome radioresistance in lung cancer therapy.

Production Conditions and Characterization of the Exo-biopolymer Produced by Submerged Cultivation of Ganoderma lucijum Mycelium (영지(Ganoderma lucidum) 균사체의 액체배양에 의한 세포외 생물고분자의 생산조건과 특성)

  • Lee, Shin-Young;Kang, Tae-Su
    • Microbiology and Biotechnology Letters
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    • v.24 no.1
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    • pp.111-118
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    • 1996
  • For the screening and the development of the new bio-material, cultural conditions for the exo-biopolymer (EBP) production throught the submerged cultivation of Ganoderma lucidum mycelium were investigated. Also, the fractionations and the purifications of the exo-biopolymer were carried out and the chemical compositions of the exo-biopolymer were examined. The optimal culture conditions for the exo-biopolymer production were pH 5.0, 30$^{\circ}C$ and 100 rpm of agitation speed in the medium containing of 5% (w/v) glucose, 0.5%(w/v) yeast extract, 0.1% (w/v) ($(NH_4)_2HPO_4$, and 0.05% (w/v) $KH_2PO_4$. In the flask cultivation for 7 days under these conditions, the concentration of the maximum exo-biopolymer and the cell mass were 15.4g/l and 18.8g/l, respectively. The specific growth rate was 0.039 $hr^{-1}$. In addition, the substrate consumption rate, and the exo-biopolymer production rate were 0.043$gg^{-1}$$hr^{-1}$ and 0.025$gg^{-1}$$hr^{-1}$, respectively. The exo-biopolymer was fractionated into BWS (water soluble exo-biopolymer) and BWI (water insoluble exo-biopolymer) by the water extraction, and the sugar contents of two fractions were higher than 97% (based on dry basis). The components sugar of BWS and BWI fractions were glucose, galactose, mannose, xylose, and fucose. Their molar ratios were 3.6:1.5:2.1:0.5: trace and 2.9:3.1:2.0:1.6:0.3, respectively.

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