• Title/Summary/Keyword: C6 세포

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Profile of Gene Expression Changes During Doxorubicin Induced Apoptosis of Saos-2 (Saos-2 세포에서 Doxorubicin에 의한 세포사멸 유도과정에서의 유전자 발현 변화)

  • Lim, Jeong-Sook;Bae, Min-Jae;Baek, Suk-Hwan;Kim, Jae-Ryong;Kim, Jung-Hye;Kim, Seong-Yong
    • Journal of Yeungnam Medical Science
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    • v.22 no.2
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    • pp.221-240
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    • 2005
  • Background: Doxorubicin has proved to be a useful chemotherapeutic agent especially for osteogenic sarcoma. It induces cancer cell death via apoptosis. Materials and Methods: To explore and analyze the changes of gene expression during doxorubicin induced apoptosis on human osteogenic sarcoma, Saos-2 cell, cDNA microarray was performed. After treatment with doxorubicin, total RNA was purified and expressed genes were investigated with a 17k human cDNA microarray. Results: For analysis of the cDNA microarray, the genes were filtered using the sum of the median value of Cy3 and Cy5 signal intensity of greater than 800. Expression of 264 genes was changed by more than 2 fold, and the expression of 35 genes was changed more than 3 fold after treatment with doxorubicin. The genes were primarily related to cell death, cell growth and maintenance, signal transduction, cellular component, transport, and metabolism. Conclusion: Treatment with doxorubicin induced expressional change of many genes. Some of the genes might be related with apoptosis directly or indirectly. Further study is now needed to characterize these genes.

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Effect of Temperature on Mitotic Cycle of Rice Root Meristem Cells (벼 뿌리세포의 유사분열주기에 대한 배양온도의 영향)

  • 김재철;이승준;권성환;곽성희
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.35 no.1
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    • pp.65-72
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    • 1990
  • The mitotic cycle duration and component plase periods of rice (Oryza sativa L. 'Sumjinbyeo and Seokwangbyeo') root merisem cells at 15, 20, and 30$^{\circ}C$ were determined the use of tritiated thymidine, In this work, the time interval between the maxima of sequential mitotic appearances of marked cells was used to estimate the mitotic cycle duration (MCD) of rice, The MCD of rice of the cultivar 'Sumjinbyeo' and 'Seokwangbyeo' at 20. and 30$^{\circ}C$ was 12. and 20hr, respectively. But the MCD of 'Sumjinbyeo' and 'Seokwangbyeo' was 18 and 20hr, at 15$^{\circ}C$. respectively. The MCD decreased with increasing temperature, The duration of component phase of rice cultivar 'Seokwangbyeo' were essentially the same ratio at 20$^{\circ}C$ and 30$^{\circ}C$. but in 'Sumjinbyeo' cultivar the ratio of $G_1$ period was almost doubled while those of $G_2$ and M were decreased by almost two times at 20$^{\circ}C$ and 30$^{\circ}C$. Deoxyribonucleic acid (DNA). Ribonucleic acid (RNA), and protein synthesis were reduced with increasing temperature from 15$^{\circ}C$ to 30$^{\circ}C$ while the MCD was decreased, This result suggest that DNA, RNA and protein synthesis may not affect the MCD from 15$^{\circ}C$ to 30$^{\circ}C$ in rice.

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IDENTIFICATION OF GENES INVOLVED IN OSTEOCLAST DIFFERENTIATION BY CDNA ARRAY ANALYSES (dDNA array를 이용한 파골세포 분화 관련 유전자의 탐색)

  • Cho, Young-Jun;Lee, Zang-Hee;Lee, Chang-Seop;Lee, Sang-Ho
    • Journal of the korean academy of Pediatric Dentistry
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    • v.29 no.2
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    • pp.278-284
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    • 2002
  • To examine the global gene expression of osteoclastogenesis-related genes in RAW 264.7 and its differentiated OCLs through the use of Atlas Mouse cDNA Array 2.1 membranes printed with 1176 well-characterized mouse genes involved in biology. Both samples were screened in parallel using cDNA expression arrays. The array results were additionally validated using RT-PCR. The results of cDNA arrays showed that 6 genes were up-regulated >2.5-fold (PKC beta II. POMC, PTEN, etc) and 16 genes were down-regulated >2.5-fold (Osteopontin, Cyclin D1, Cathepsin C, PTMA, etc) in both samples at the mRNA level. RT-PCR analysis of PKC beta II of these differentially expressed genes gave result consistent with cDNA array findings. The result of osteoclastogenesis showed that the PKC beta II gene was overexpressed in OCLs compared with RAW264.7 cell line. Osteoclastogenesis-related genes are differentially expressed in RAW264.7 cell line and its differentiated OCLs. its gene overexpression correlates with osteoclast differentiation in RAW264.7 cell line.

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The Effects of the Expression of GATA Binding Protein 6 on Heart and Brain Development (심장과 뇌 발달에서 GATA6 유전자 발현 감소가 미치는 영향)

  • Seo, Jungwon
    • Journal of Life Science
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    • v.25 no.11
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    • pp.1230-1234
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    • 2015
  • GATA binding protein 6 (GATA6) is a transcription factor that is expressed in the early blastocyst stage and controls the expression of important genes in the differentiation and development of the heart, pancreas, and intestine. This study confirmed the role of GATA6 in cell differentiation and organ development using mouse embryonic stem cells and zebrafish, respectively. First, the mouse embryonic stem cells were differentiated into pacemaker cardiomyocytes. An RT-PCR analysis revealed that the expression of the GATA6 gene was greatly increased from day 4 of differentiation. The expression of GATA6 was upregulated prior to increased expression of NK2 homeobox 5 (Nkx2.5) and myocyte enhancer factor 2C (MEF2C), which are critical transcription factors involved in regulating heart formation. To examine the role of GATA6 in development, GATA6 morpholino was microinjected into zebrafish embryos. Knockdown of GATA6 expression significantly decreased the heart size and heart rate in the zebrafish compared to a control. In addition, the brains were degenerated in the GATA6 morpholino-injected zebrafish. Acridine orange staining showed that knockdown of GATA6 expression increased apoptotic cells in the brain. Interestingly, knockdown of GATA6 expression decreased apoptotic cells in the early bud stage. This study points to the importance of the GATA6 gene in heart and brain development.

Studies on the Efficient Embryo Transfer Methods using Inbred Embryos in Generation of Transgenic Mice (Inbred 마우스 수정란을 이용한 형질전환마우스 생산에 있어서의 효과적인 수정란 이식 방법에 관한 연구)

  • Wang A. G;Kim, S. U.;Y. H. Han;Kim, S. K.;D. Y. Yu
    • Korean Journal of Animal Reproduction
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    • v.27 no.1
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    • pp.69-75
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    • 2003
  • The objective of this study is to improve the efficiency of embryo transfer in generation of transgenic mice using inbred mouse (C57BL/6J) embryos. The embryos of C57BL/6J and BCF1 mice were superovulated by the standard protocol. One-cell stage of embryos were microinjected and the resulted one-or two-cell were transferred into one-or two-side oviducts of foster mother, respectively. When one-cell stage of embryos were transferred into one-side oviduct of 0.75 d.p.c. foster mother, the results were not ideal because of showing pregnancy ratios of 68.8$\pm$7.83% for C57BL/6J and 48.3$\pm$14.22% for BCF1, and development ratios of pups vs transferred embryos of 11.9$\pm$5.51% for C57BL/6J and 10.5$\pm$8.03% fur BCF1 . However, when two-cell stage of embryos were transferred into two-side oviducts of 0.5 d.p.c. foster mother, we got better results of 94.4$\pm$9.64% and 100$\pm$0% pregnancy ratio, and 22.1$\pm$0.4% and 21.8$\pm$0.38% development ratio for C57BL/6J and BCF1, respectively. Therefore, transferring two-cell stage of C57BL/6J embryos into two-side oviducts of 0.5 d.p.c. foster mother may be competitive to the result in BCF1 embryos. Even if there are a lot of other factors affecting these results, we conclude from these experiments that transfer of two-cell embryos into two-side oviducts of 0.5 d.p.c. foster mother is a more efficient and safe method than others in generating transgenic mice using inbred mice embryos.

Effect of Resveratrol on the Induction of Cdk Inhibitor p21 and Pro-apoptotic Bax Expression by amyloid-β in Astroglioma C6 Cells (신경교 세포에서 resveratrol이 amyloid-β에 의해 유도되는 Cdk inhibitor p21 및 Bax 발현의 감소 효과)

  • Kim Young Ae;Lim Sun-Young;Ko Woo Shin;Choi Byung Tae;Lee Yong Tae;Rhee Sook-Hee;Park Kun-Young;Lee Won-Ho;Choi Yung Hyun
    • Journal of Life Science
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    • v.15 no.2 s.69
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    • pp.169-175
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    • 2005
  • Resveratrol (3,4',5-trihydroxy-trans-stilbene), a phytoalexin found in grape skins, peanuts, and red wine, has been reported to have a wide range of biological and pharmacological properties. $Amyloid-\beta$ deposition and senile plaque-associated astrocytes are common neuropathological features of Alzheimer's disease. In this study, we have explored the effects of resveratrol on $amyloid-\beta-peptide-mediated$ cytotoxicity in vitro and modulation of cell growth-regulatory gene products in astroglioma C6 cells to elucidate its possible mechanism for anti-cytotoxicity. Exposure of C6 cells to $Amyloid-\beta$ resulted in dose-dependent growth inhibition and morphological changes of C6 cells, which were recovered by pre-treatment with resveratrol. The anti-proliferative effect of $amyloid-\beta$ was associated with the induction of tumor suppressor p53 and cyclin-dependent kinase (Cdk) inhibitor p21 (WAF1/CIP1) expression assessed by RT-PCR and Western blot analysis in time-dependent manner in C6 cells. In addition, the pro-apoptotic Bax expression was also up-regulated in $amyloid-\beta-treated$ C6 cells without alteration of anti-apoptotic Bcl-2 and $Bcl-X_L$ expression. However, pre-treatment of resveratrol significantly inhibited $amyloid-\beta-induced$ p53, p21 and Bax levels, suggesting that the modulation of p53, p21 and Bax levels could be one of the possible pathways by which resveratrol functions as anti-cytotoxic agent. Our results demonstrate that resveratrol may enhance the protection against $amyloid-\beta-induced$ cytotoxicity by promoting the survival of glial cells.

Effects of Mercury Chloride on Nitric Oxide Syntheses in Mouse Peritoneal Macrophage and EMT-6 Cell (Mercury Chloride가 마우스 복강대식세포 및 EMT-6 세포의 Nitric Oxide 생성에 미치는 영향)

  • Kwon, Keun-Sang;Koh, Dai-Ha;Ki, No-Suk;Youm, Jung-Ho
    • Journal of Preventive Medicine and Public Health
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    • v.30 no.2 s.57
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    • pp.369-380
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    • 1997
  • The effects of treatment with mercury chloride on the nitrite and nitrate syntheses were observed in peritoneal macrophages from Balb/c mice and EMT-6 cells in vitro. The cells were cultured in Dulbecco's modified Eagle's medium(DMEM) with cytokines. Amounts of nitrite and nitrate in the culture media after 24 and 36 hours of culture were about 2-fold, and 3-fold of those measured after 12 hours respectively. There were very close associations Between the amounts of nitrite and nitrate measured in the culture media according to culture time. The survival rate of peritoneal macrophages was significantly decreased by mercury chloride added into the media in dose-dependent manner, however the survivals of EMT-6 cells were not influenced by mercury chloride concentration in media. Nitrite and nitrate syntheses were dose-dependently decreased by mercury chloride added in culture media. ATP synthesis also decreased in EMT-6 cells by mercury chloride. These results reported here suggest that the disorder of cell mediated immunity by mercurials could be related to the inhibition of nitric oxide synthesis which seems to be caused by the inhibition of ATP synthesis.

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Cyanate Induces Apoptosis of Rat Glioma Cell Line (시안산에 의한 신경아교종세포의 자멸사)

  • Choi, Hye-Jung;Lee, Sang-Hee
    • 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.

Free radical scavenging activity and protective effect of three glycyrrhiza varieties against hydrogen peroxide-induced oxidative stress in C6 glial cells (종류별 감초의 라디칼 소거능 및 H2O2에 의한 C6 glial 세포의 산화적 스트레스 개선 효과)

  • Kim, Ji Hyun;Cho, Min Ji;Park, Chan Hum;Cho, Eun Ju;Kim, Hyun Young
    • Journal of Applied Biological Chemistry
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    • v.63 no.4
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    • pp.327-334
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    • 2020
  • Oxidative stress is common cause of neurodegenerative diseases. The purpose of this study is to investigate the in vitro free radical scavenging activity and protective effect of three Glycyrrhiza species including Glycyrrhiza uralensis, G. glabra, and a new variety of Glycyrrihza (Shinwongam, SW) against hydrogen peroxide-induced oxidative stress in C6 glial cells. In vitro assays, radical scavenging activities of G. uralensis, G. glabra, and SW against 2,2-diphenyl-1-picrylhydrazyl, ·OH, and O2- increased as concentration-dependent manner. In addition, the SW was found to contain the highest polyphenol and flavonoid contents. The treatment of H2O2 to C6 glial cell induced oxidative stress, whereas G. uralensis, G. glabra, and SW significantly increased the cell viability as dose-dependent manner. In particular, SW exerted stronger protective effect on H2O2-induced cytotoxicity, than G. uralensis and G. glabra. Furthermore, reactive oxygen species (ROS) formation was significantly elevated by H2O2 in C6 glial cells. However, treatments of G. uralensis, G. glabra, and SW decreased ROS formation. In addition, SW decreased pro-inflammatory related protein expression levels such as inducible nitric oxide synthase and cyclooxygenase-2, compared to H2O2-treated control group. These results indicated that G. uralensis and G. glavra, especially SW, may be useful for preventing from oxidative stress-induced neuronal damage by regulating inflammatory reaction.

Anti-Obesity Effect of Ethyl Acetate Fraction from 50% Ethanol Extract of Fermented Curcuma longa L. in 3T3-L1 Cells (발효울금 주정추출물부터 분리된 에틸아세테이트 분획물에 대한 3T3-L1 세포에서의 지방 형성 억제 효과)

  • Kim, Jihye;Park, Jeongjin;Jun, Woojin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.11
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    • pp.1681-1687
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    • 2014
  • In the present study, we investigated the effect of ethyl acetate fraction from 50% ethanol extract of fermented Curcuma longa L. (FCEE) on lipid metabolism in 3T3-L1 cells. The safety range of FCEE was up to $300{\mu}g/mL$. Effects of FCEE on lipid accumulation and intracellular triglyceride (TG) content in 3T3-L1 cells were examined by Oil Red O staining and AdipoRed assay. Compared to adipocytes, lipid accumulation and intracellular TG content were significantly reduced by 10.2% and 13.7%, respectively, upon FCEE treatment at a concentration of $200{\mu}g/mL$. Glucose uptake by 3T3-L1 cells was significantly reduced by 36.6% compared to adipocytes at a concentration of $200{\mu}g/mL$. On day 8, free glycerol release into the culture medium was significantly reduced compared to adipocytes at concentrations of 50, 100, and $200{\mu}g/mL$ of FCEE. FCEE significantly stimulated RNA expression of AMP-activated protein kinase (AMPK) and suppressed mRNA expressions of sterol regulatory element-binding protein-1c (SREBP-1c), CCAAT/enhancer binding proteins ${\alpha}$ ($C/EBP{\alpha}$), and peroxisome proliferator- activated receptor ${\gamma}$ ($PPAR{\gamma}$) in 3T3-L1 cells. These results suggest that FCEE inhibits adipogenesis through activation of AMPK mRNA expressions and inhibition of SREBP-1c, $C/EBP{\alpha}$, and $PPAR{\gamma}$ mRNA expressions.