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Effects of Inhibitors on the Function and Activity of Topoisomerase, and Gene Expression in HL-60 Human Leukemia Cells (HL-60 세포의 유전자 발현 및 topoisomerase의 기능 활성에 미치는 억제제의 영향)

  • Jeong, In-Cheol;Cho, Moo-Youn;Park, Jang-Su
    • Journal of Life Science
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    • v.18 no.1
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    • pp.75-83
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    • 2008
  • This studies were designed to elucidate whether inhibitors of topoisomerase regulate function and activity of topoisomerase, and gene expression in HL-60 human leukemia cells. HL-60 cells were treated with 10-hydroxycamptothecin or doxorubicin, total RNA was isolated, and expressed genes were investigated with human oligonucleotide microarray containing 10K gene, respectively. Expression profiles of the human leukemia HL-60 cells treated with 10-hydroxycamptothecin (10-CIT) or doxorubicin associated with signal transduction,. cell adhesion, cell cycle, cell growth, cell proliferation, cell differentiation, transcription and immune response, especially genes related with transcription and cell growth. In HL-60 cells treated with 10-CPT, the expression of topoisomerase III${\alpha}$, III${\beta}$ and I gene from oligo chip microarray analysis were increased over, but the expression of topoisomerase II${\alpha}$ and II${\beta}$ gene were decreased over. In contrast, the expression of topoisomerase II${\alpha}$ and II${\beta}$ gene were increased over in HL-60 cells treated with doxorubicin, whereas the expression of topoisomerase III${\alpha}$ and III${\beta}$ mRNA remained no significant change. These results suggest that these data may be useful for novel therapeutic markers.

Correlation of Gene Expression between Adiponectin and Glucose Transporter 4 in Mouse Adipose Tissue (생쥐 지방조직에서의 아디포넥틴과 포도당수송체-4 유전자 발현의 상관관계)

  • Lee, Yong-Ho
    • Journal of Life Science
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    • v.24 no.8
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    • pp.895-902
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    • 2014
  • Adiponectin has been known to improve insulin sensitivity and elicit glucose uptake via increased glucose transporter 4 (GLUT4) translocation. In the current study, mRNA expression levels of adiponectin and GLUT4 were measured in subcutaneous adipose tissue from C57BL/6 mice fed normal (ND) or high-fat diet (HFD) until 16, 26, 36, 47, or 77 weeks of age starting from 6 weeks of age. Expression levels were also measured in mice with calorie restriction (CR) and in thiazolidinedione (TZD) treated mice. Using quantitative real-time PCR, we demonstrated that GLUT4 expression in adipose tissue significantly decreased in HFD mice groups and increased in CR (p<0.05) and TZD (p=0.007) groups while there was no difference in adiponectin mRNA expression levels between experimental and control groups. General linear regression models were used to assess the association of gene expression levels between adiponectin and GLUT4 and to determine whether adiponectin affects GLUT4 transcription. mRNA expression levels of adiponectin and GLUT4 are significantly associated each other in mice fed a ND (p<0.0001) or HFD (p<0.0001), in groups separated into each age and diet, and CR group (p=0.002), but not in TZD group (p=0.73). These results demonstrated that gene expression of adiponectin and GLUT4 is strongly associated, suggesting that there is a common regulatory mechanism for adiponectin and GLUT4 gene expression and/or adiponectin has a direct role in GLUT4 gene expression in adipose tissue.

Induction of G1 Phase Cell Cycle Arrest and Apoptotic Cell Death by 5-Fluorouracil in Ewing′s Sarcoma CHP-100 Cells (CHP-100 Ewing′s 육종세포에서 5-fluorouracil에 의한 G1 arrest 유도 및 apoptosis 유발에 관한 연구)

  • Kim, Sung Ok;Choi, Yung Hyun
    • Journal of Life Science
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    • v.26 no.9
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    • pp.1015-1021
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    • 2016
  • 5-fluorouracil (5-FU), a pyrimidine analog, is a widely used anticancer drug, which works through irreversible inhibition of thymidylate synthase. In the present study, it was investigated the anti-proliferative effects and molecular mechanisms of 5-FU using Ewing's Sarcoma CHP-100 Cells. The present data indicated that treatment of 5-FU to CHP-100 cells induced a G1 phase arrest of the cell cycle in a time-dependent manner. 5-FU-induced G1 arrest was correlated with the accumulation of the hypophosphorylated form of the retinoblastoma protein (pRB) and association of pRB with the transcription factors E2F-1 and E2F-4. Although 5-FU treatment did affect the levels of cyclin-dependent kinases, the levels of cyclin A and B were markedly down-regulated as compared with the untreated control group. In addition, 5-FU-induced G1 arrest of CHP-100 cells was also associated with the induction of apoptosis, as determined by apoptotic cell morphologies, degradation of poly(ADP-ribose) polymerase and Annexin V staining. Furthermore, 5-FU induced the loss of mitochondrial membrane potential with up-regulated pro-apoptotic Bax expression, down-regulated anti-apoptotic Bcl-2 expression and cytochrome c release from mitochondria to cytosol. Collectively, the data suggest that 5-FU is effective in inducing cell growth reduction and apoptosis, in part, by reducing phosphorylation of pRB and activating mitochondrial dysfunction in CHP-100 cells.

SREBP as a Global Regulator for Lipid Metabolism (지질대사 조절에서 SREBP의 역할)

  • Lee, Wonhwa;Seo, Young-kyo
    • Journal of Life Science
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    • v.28 no.10
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    • pp.1233-1243
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    • 2018
  • Sterol regulatory-element binding proteins (SREBPs) are a family of transcription factors that regulate lipid homeostasis and metabolism by controlling the expression of enzymes required for endogenous cholesterol, fatty acid (FA), triacylglycerol, and phospholipid synthesis. The three SREBPs are encoded by two different genes. The SREBP1 gene gives rise to SREBP-1a and SREBP-1c, which are derived from utilization of alternate promoters that yield transcripts in which distinct first exons are spliced to a common second exon. SREBP-2 is derived from a separate gene. Additionally, SREBPs are implicated in numerous pathogenic processes, such as endoplasmic reticulum stress, inflammation, autophagy, and apoptosis. They also contribute to obesity, dyslipidemia, diabetes mellitus, and nonalcoholic fatty liver diseases. Genome-wide analyses have revealed that these versatile transcription factors act as important nodes of biological signaling networks. Changes in cell metabolism and growth are reciprocally linked through SREBPs. Anabolic and growth signaling pathways branch off and connect to multiple steps of SREBP activation and form complex regulatory networks. SREBPs are activated through the PI3K-Akt-mTOR pathway in these processes, but the molecular mechanism remains to be understood. This review aims to provide a comprehensive understanding of the role of SREBPs in physiology and pathophysiology at the cell, organ, and organism levels.

Cloning and Characterization of Homeodomain-Zip Gene, Phc5 in Embryogenic Callus derived from Pimpinella brachycarpa Suspension Cultured Cells (참나물 현탁배양세포 유래 배발생캘러스에서 HD-Zip 유전자, Phc5의 클로닝과 특성)

  • 손수인;김준철
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.2
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    • pp.121-126
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    • 1999
  • Calli were induced from the petiole explants of Pimpinella brachycarpa on MS medium supplemented with 0.5 mg/L 2,4-D and 0.1 mg/L BA after four weeks of culture. Compact clusters of small and dense cells among these calli were selected and suspension-cultured as the source of embryogenic calli. When transferred to MS medium with 0.1 mg/L NAA, the suspension-cultured cells grew to embryogenic callus. Somatic embryos derived from these embryogenic calli developed into plantlets. The cDNA library was constructed in the embryogenic callus and in order to screen the cDNA library, these cDNAs were plated at a density 1.5 $\times$ 10^5 plaques per 15 cm petridish. Among 19 clones showing preferential hybridization with petiole HD-Zip gene, five clones were obtained after second screening. Four clones among them, were highly homologous to P. brachycarpa shoot-tip Phz4 gene, but one clone, Phc5 was about 1.5 kb which has an extra 163 bp to 5' upstream of Phz4. The Phc5 was 1,531 bp containing poly A tails of 18 bases. ATG start codon for Phc5, was located at position 284 with an open reading frame of 906 by which encodes a polypeptide of 302 amino acids. The Phc5 protein revealed that the polypeptides between 135 and 195 contain a homeodomain as the `leucine zipper' motif.

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A Study On Characteristics Of Forged Sealing Made Of Photosensitive Resin (감광성 수지로 제작된 위조 인영의 특성에 관한 연구)

  • Han, Sang-Deog;Jeong, Yang-Kwon;Choi, Jae-Ho
    • The Journal of the Korea institute of electronic communication sciences
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    • v.7 no.6
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    • pp.1385-1391
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    • 2012
  • The authenticity of a document is recognized only when sealing on the document can be recognized to be the same as the original seal of a person who drew up the document. But forge in various ways is prevalent; forge through stamp-copy, forge through photocopying, forge through computer seal sculpture machine, forge through photosensitive resin, etc. Among the methods above, a method of making a forged seal with photosensitive resin panels, which is one of the easiest forging methods, enables making a forged seal the same as the original seal through the work of projecting ultraviolet rays on a resin panel on which negative film is stuck after making it using the original seal. But as research into this reality has not been sufficiently carried on, it is difficult to identify forged sealing through photosensitive resin seals, indeed. For this reason, unless regulations about making forged seals using resin machine for print are enforced, crimes related to forged seals using photosensitive resin are expected to increase in the future. Thus, after stamping 10 resin seals made based on an original seal carved of thick and thin strokes of characters, this paper intends to clarify that there are differences between original sealing and forged sealing through sealing identification method. Consequently, we verified the significant difference in the statistics between original sealing and forged sealing.

3 Tesla MR Clinical Application: Advanced Neuroimaging

  • 손철호
    • Proceedings of the KSMRM Conference
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    • 2002.11a
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    • pp.50-56
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    • 2002
  • 최근 3-4년간 MR의 hardware와 software의 급격한 발전으로 마침내 작년 말에 3.0 Tesla whole body MR 장비가 미국 FDA의 공인을 받았다. 한국에서도 일찍부터 3T MR장비의 개발이 이루어 졌고 이미 설치되어 연구와 임상이 이용되고 있다. 여러 회사에서 개발 및 연구된 전신 3.0T MR 장비가 여러가지 가능성을 보이고 임상 도입 단계에 있지만 아직까지 실지 임상에서는 뇌신경계 분야가 주류를 이루고 있다. 지금 뇌신경계 분야에서 보편적으로 늘리 사용되고 있는 1.5T MR 장비는 모든 면에서 상당히 안정적으로 임상 및 연구에 이용되고 있다. 1-2년 전만 해도 3.0 T MR기기는 뇌신경계 영역에서도 임상적으로 늘리 사용되기에는 안정적인 면에서는 1.5T 기기에 비해서 떨어지는 것이 사실이었다. 그래서 주로 연구실 영역에서 많이 이용되고 있었다. 그러나 지금 본원에 설치 완료되어 임상에 적용한지 6개월 정도 이용한 예에서 보면 (about 2300 cases/6months) hardware, software적인 면에서 아직 조금의 불편함이 있지만 많은 부분이 충분히 인지되고 개선이 가능한 부분으로 거의 불편함이 사라질 것으로 기대되고 있고, 불편함을 넘을 수 있는 여러 가지 장점이 있다고 본다. 고자장 (>3.0 T) MRI의 매력은 자장에 비례적으로 SNR, spectral resolution이 높아지고, T1, BOLD등에 의한 대조도가 향상한다는 것이다. SNR의 증가는 temporal, spatial 분해능을 증가시키고, spectral resolution이 높아짐에 따라 MR spectroscopy상에서 주요 대사물질 이외 작은 대사물질에 관한 스펙트럼의 분석을 향상시킨다. 이처럼 고자장 MR은 근본적인 장점을 가지고 있고 이러한 장점이 고자장 MR 시대로 가야 할 이유을 모두 설명하고 있다고 생각된다.세포질등이 있으며, 이들중에서 lysosomes, peroxisomes, 그리고 미토콘드리아가 특정한 유전성 백질질환에 중요한 역할을 하는 것이 밝혀졌다. 이러한 질환들은 최소한 각 소기관에 의한 질환군으로 분류될 수 있다.SXR이 ER의 transactivation 효과를 약간 촉진한 반면 MDA-MB-231세포는 SXR을 제외한 CAR와 PPAR${\gamma}$에 의해 ER의 transactivation 효과가 약간 증가되는 경향을 보였다. 이러한 결과는 유방암세포에서는 CAR, SXR, PPAR${\gamma}$과 같은 xenobiotic nuclear receptor에 의한 ER transactivation 효과가 간암세포와는 다르게 나타나며, 유방암의 종류에 따라서 endogenous CAR, SXR, PPAR${\gamma}$수용체가 다르게 발현됨으로써 이들에 대한 반응이 서로 상이한 특징을 나타낼 수 있을 것으로 사료된다. 따라서 estrogen receptor에 의해 매개되는 estrogn의 전사활성조절기전이 표적세포에 따라 다른 경로를 포함 할 수 있음을 시사한다.서 흡착 능력이 우수하게 나타났으며, 황화수소는 펄라이트, 왕겨, 소나무수피에서 상대적으로 우수한 것으로 나타났으며, 혼합충전재는 암모니아의 경우 코코넛과 펄라이트의 비율이 7:3인 혼합 재료 3번과 소나무수피와 펄라이트의 비율이 7:3인 혼합 재료 6번에서 다른 혼합 재료에 비하여 우수한 것으로 나타났다. 4. 코코넛과 소나무수피의 경우 암모니아 가스에 대한 흡착 능력은 거의 비슷한 것으로 사료되며, 코코넛의 경우 전량을 수입에 의존하고 있다는 점에서 국내 조달이 용이하며, 구입 비용도 적게 소요되는 소나무수피를 사용하는 것이 경제적이라고 사료된다. 5. 마지막으로 악취제거 미생물균주를 접종한 소나무수피 50%와 펄라이트

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Expression of Fra1 and Fra2 Genes are regulated by Estrogen in the Mouse Uterus (생쥐자궁에서 에스트로겐에 의해 조절되는 Fra1과 Fra2 유전자의 발현양상)

  • Lee, Ji-Yoon;Hong, Seok-Ho;Nah, Hee-Young;Kim, Sung-Hoon;Chae, Hee-Dong;Kim, Chung-Hoon;Kang, Byung-Moon;Kim, Moon-Kyoo
    • Clinical and Experimental Reproductive Medicine
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    • v.30 no.4
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    • pp.309-316
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    • 2003
  • 연구목적: 스테로이드 계통의 에스트로겐 호르몬은 막 수용체와 결합하고 DNA에 부착되어, 자궁조직에서 발현되는 많은 유전자들의 발현 양상을 조절하는 것으로 알려져 있다. 본 연구에서는 난소를 제거한 생쥐 모델을 이용하여 에스트로겐에 의해 조절되는 전사 관련 유전자(transcription factor)들을 동정하고, early up-regulation gene으로 확인된 Fos related antigen (Fra1과 Fra2) 유전자의 발현 양상을 RT-PCR과 면역염색방법으로 살펴보았다. 연구재료 및 방법: 난소 절제술을 시행한 생쥐에 에스트로겐을 피하주사하고 2, 4, 6, 12시간 간격으로 자궁조직을 적출하였다. 대조군으로는 sesame oil만을 주사한 후 2시간째에 수획한 자궁조직을 사용하였으며, 시간대별로 채취한 자궁조직(n=4)에서 RT-PCR을 수행하였다. RT-PCR을 통해 early response gene으로 확인된 Fra1과 Fra2에 대한 에스트로겐의 영향을 살펴보기 위해 estrogen receptor antagonist인 ICI 182, 780을 주사하여 유전자 발현 양상의 변화를 살펴보았다. 또한, 자궁조직내에서의 단백질 발현 부위를 관찰하기 위해 면역조직화학염색을 실시하였다. 결 과: 생쥐 자궁조직에서 에스트로겐에 의해 발현 양상의 변화가 확인된 유전자는 early up-regulation genes (CREB2, Fra-1, 2, GATA5), late up-regulation gene (E2F1), no response genes (CREB1, ATF1, GLI3, E2F3), down-regulation genes (GLI2, E2F5, GATA-2, 3, 6) 등으로 구분할 수 있었다. 그 중 early up-regulation genes에 해당하는 Fra1과 Fra2 유전자는 ICI 182, 780에 의해 그 발현이 유의하게 감소되는 것을 확인하였다(p < 0.01). 이들 단백질은 생쥐 자궁조직의 상피세포층, 기질층, 근육층에서 고루 발현되었으며, 특히 근육층에서 강한 염색정도를 관찰할 수 있었다. 결 론: 이상의 결과를 통해 Fra1과 Fra2 유전자의 발현은 에스트로겐에 의해 조절됨을 알 수 있었으며, 이들의 강한 발현이 자궁조직의 근육층에서 관찰되어 이들의 기능에 대한 연구가 필요할 것으로 생각된다.

Phosphorylation Properties of Recombinant OsCPK11, a Calcium-dependent Protein Kinase from Rice (벼의 칼슘-의존적 단백질 카이네즈인 재조합 OsCPK11의 인산화 특성)

  • Cho, Il-Sang;Lee, Su-Hee;Park, Chung-Mo;Kim, Sung-Ha
    • Journal of Life Science
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    • v.27 no.12
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    • pp.1393-1402
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    • 2017
  • In plants, calcium ($Ca^{2+}$)-dependent protein kinases (CDPKs) are important sensors of $Ca^{2+}$ signals. Previous research demonstrated the expression of the OsCPK11 gene in various tissues at the transcription level, but its developmental and biochemical functions at the protein level were not determined. This study was aimed to identify biochemical characteristics of OsCPK11. GST- OsCPK11 was expressed in E. coli and used for an in vitro kinase assay. Biochemical analyses identified OsCPK11 as a CDPK. OsCPK11 autophosphorylated itself and transphosphorylated histone III-s and MBP as substrates in the presence of $Ca^{2+}$. The activity of the recombinant OsCPK11 was influenced by $Mg^{2+}$, with optimum activity detected at pH 7.0-7.5. OsCPK11 activity was not affected by $Mg^{2+}$, $Mn^{2+}$, or $Na^+$ in the presence of a high level of $Ca^{2+}$. Autophosphorylation of OsCPK11 decreased $Ca^{2+}$ sensitivity of OsCPK11. An anti-OsCPK11 rabbit antibody recognized 95.5 kD of GST-OsCPK11, as shown by an immunoblot analysis. These results shed light on the function of OsCPK11 in $Ca^{2+}$-mediated signaling in rice.

Gene Structure and Function of fkhE, a Forkhead Gene in a Filamentous Fungus Aspergillus nidulans (Aspergillus nidulans forkhead 유전자 fkhE의 구조와 기능 분석)

  • Park, Mi-Hye;Kim, Hyoun-Young;Kim, Jong-Hwa;Han, Kap-Hoon
    • The Korean Journal of Mycology
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    • v.38 no.2
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    • pp.160-166
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    • 2010
  • A homothallic filamentous fungus Aspergillus nidulans has been used as the a model organism for studying growth and development for eukaryotic system. Various studies about specific transcription factors have been performed for elucidating the molecular mechanisms of growth, asexual and sexual developmental processes. Among them, the fkhE gene (AN2025.3) is located in chromosome VII and contains an ORF encoding 718 amino acid polypeptide intervening with two short introns. The cDNA sequencing revealed that at least four types of alternative splicing events were occurred when the fkhE gene was transcribed. The putative FkhE polypeptide contains a conserved forkhead domain and a bipartite nuclear localization signal at it's N-terminus and C-terminus, respectively. Deletion of fkhE resulted in impaired conidiophore formation in a solid medium. However, the sexual developmental process or cleistothecia formation was normal. Furthermore, fkhE deletion mutant produced conidiophores and conidia under the submerged culture, indicating that the fkhE gene is involved in asexual developmental process similar to the fkhF gene.