• Title/Summary/Keyword: CPT-1

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Induction of Apoptosis by Camptothecin in HL-60 Cells (HL-60 세포에서 Camptothecin의 apoptosis 유도작용)

  • 김해종;천영진;김미영
    • YAKHAK HOEJI
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    • v.43 no.3
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    • pp.385-390
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    • 1999
  • Camptothecin (CPT) has been known to induce apoptosis in various cancer cell lines. To examine the intracellular apoptotic death signal initiated by CPT, we investigated the possible connection between caspase-3 activation and GSH depletion during CPT-induced apoptosis in HL-60 cells. Treatment of cells with $1{\;}{\mu}M$ CPT induced PARP cleavage accompanied by DNA fragmentation. z-VAD-fmk, a caspase-3 inhibitor, blocked the CPT-induced DNA fragmentation. Pretreatment of cells with N-acetylcysteine, a precursor of GSH biosynthesis, failed to inhibit CPT-induced PARP celavage and DNA gragmenatation. No significant changes in GSH depletion is not essential for caspase activation during CPT-induced apoptosis. We also investigated whether CPT-induced apoptosis is associated with changes of the levels of Bax and Bcl-2, two proteins involved in the control of apoptosis. Bcl-2 levels exhibited a late decrease compared with the kinetics of DNA fragmentation, whereas Bax levels increased more rapidly after CPT treatment. These results suggest that Bax plays more important role than Bcl-2 in inducing DNA fragmentation and may function upsteam of proteolytic activation of caspase-3 pathway in CPT-induced apoptosis.

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Effects of camptothecin on the expression of DNA topoisomerase I and c-myc in HL-60 human leukemia cells (HL-60 사람 백혈병 세포에서 camptothecin이 DNA topoisomerase l과 c-myc의 발현에 미치는 영향)

  • 정인철;정대성;류경자;박장수;조무연
    • Journal of Life Science
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    • v.10 no.6
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    • pp.621-629
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    • 2000
  • Camptothecin (CPT) is an antitumor alkaloid that has been isolated from the Chinese tree, Camptotheca acuminata. The cytotoxicity of CPT has been correlated to its inhibition of DNA topoisomerase (Topo) I by stabilizing drug-enzyme-DNA “cleavable complex" resulting in DNA single-strand breaks and DNA-protein crosslinks. This studies were designed to elucidate whether CPT regulates Topo I mediated by CPT in DNAs containing c-myc protooncogene. We have conducted experiments on Topo I purification, pUC-MYC I cloning and Topo I assay using electrophoresis, quantitative RT-PCR and Northern blotting techniques. CPT ingibited the relaxation activity of Topo I in pUC19 DNA at various concentrations (1-1000 $\mu$M), while it enhanced the cleavage of Topo I in the pUC-MYC I by forming a cleavable complex at relatively high concentrations (100-1000 $\mu$M). In HL-60 cells treated with CPT, the expression of c-myc gene was decreased over that in the control group with no changes in the expression of Topo I mRNA. Our results suggest that Topo I is the target of CPT cytotoxicity but it does not affect Topo I extression, and the suppression of c-myc mRNA expression by CPT is due to c-myc damage resulted from formation of a cleavable complex with CPT. CPT.

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cis-Prenyltransferase interacts with a Nogo-B receptor homolog for dolichol biosynthesis in Panax ginseng Meyer

  • Nguyen, Ngoc Quy;Lee, Sang-Choon;Yang, Tae-Jin;Lee, Ok Ran
    • Journal of Ginseng Research
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    • v.41 no.3
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    • pp.403-410
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    • 2017
  • Background: Prenyltransferases catalyze the sequential addition of isopentenyl diphosphate units to allylic prenyl diphosphate acceptors and are classified as either trans-prenyltransferases (TPTs) or cis-prenyltransferases (CPTs). The functions of CPTs have been well characterized in bacteria, yeast, and mammals compared to plants. The characterization of CPTs also has been less studied than TPTs. In the present study, molecular cloning and functional characterization of a CPT from a medicinal plant, Panax ginseng Mayer were addressed. Methods: Gene expression patterns of PgCPT1 were analyzed by quantitative reverse transcription polymerase chain reaction. In planta transformation was generated by floral dipping using Agrobacterium tumefaciens. Yeast transformation was performed by lithium acetate and heat-shock for $rer2{\Delta}$ complementation and yeast-two-hybrid assay. Results: The ginseng genome contains at least one family of three putative CPT genes. PgCPT1 is expressed in all organs, but more predominantly in the leaves. Overexpression of PgCPT1 did not show any plant growth defect, and its protein can complement yeast mutant $rer2{\Delta}$ via possible protein-protein interaction with PgCPTL2. Conclusion: Partial complementation of the yeast dolichol biosynthesis mutant $rer2{\Delta}$ suggested that PgCPT1 is involved in dolichol biosynthesis. Direct protein interaction between PgCPT1 and a human Nogo-B receptor homolog suggests that PgCPT1 requires an accessory component for proper function.

Evaluation of Compressibility for Normally Consolidated South-east Coast Clay Using CPT and DMT (CPT와 DMT를 이용한 남동해안 정규압밀 점토의 압축성 추정)

  • Hong, Sung-Jin;Chae, Young-Ho;Lee, Moon-Joo;Lee, Woo-Jin
    • Journal of the Korean Geotechnical Society
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    • v.27 no.4
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    • pp.21-32
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    • 2011
  • A series of in-situ and laboratory tests were performed for the clayey soils of Busan area in order to develop the methods to evaluate the compressibility using CPT and DMT results. The laboratory tests show that the clayey layers of Busan areas are normally consolidated, and their compression indices are turned out to be 0.5~1.3. From the analysis of test results, correlation factors between the cone resistance and constrained modulus (${\alpha}_m$ and ${\alpha}_n$) are observed to decrease with increasing plasticity index, and the correlation factor between the dilatometer modulus and constrained modulus $(R_M)$ increases with $1/I_D$. Based on these relationships, the methods evaluating the constrained modulus from CPT and DMT results are suggested. It is shown that the prediction method by CPT underestimates the constrained modulus of improved ground, whereas the prediction method by DMT is suitable for evaluating the constrained modulus of improved and unimproved ground.

Cryptotanshinone inhibits TNF-α-induced LOX-1 expression by suppressing reactive oxygen species (ROS) formation in endothelial cells

  • Ran, Xiaoli;Zhao, Wenwen;Li, Wenping;Shi, Jingshan;Chen, Xiuping
    • The Korean Journal of Physiology and Pharmacology
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    • v.20 no.4
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    • pp.347-355
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    • 2016
  • Cryptotanshinone (CPT) is a natural compound isolated from traditional Chinese medicine Salvia miltiorrhiza Bunge. In the present study, the regulatory effect and potential mechanisms of CPT on tumor necrosis factor alpha ($TNF-{\alpha}$) induced lectin-like receptor for oxidized low density lipoprotein (LOX-1) were investigated. Human umbilical vein endothelial cells (HUVECs) were cultured and the effect of $TNF-{\alpha}$ on LOX-1 expression at mRNA and protein levels was determined by Real-time PCR and Western blotting respectively. The formation of intracellular ROS was determined with fluorescence probe $CM-DCFH_2-DA$. The endothelial ox-LDL uptake was evaluated with DiI-ox-LDL. The effect of CPT on LOX-1 expression was also evaluated with SD rats. $TNF-{\alpha}$ induced LOX-1 expression in a dose- and time- dependent manner in endothelial cells. $TNF-{\alpha}$ induced ROS formation, phosphorylation of $NF-{\kappa}B$ p65 and ERK, and LOX-1 expression, which were suppressed by rotenone, DPI, NAC, and CPT. $NF-{\kappa}B$ inhibitor BAY11-7082 and ERK inhibitor PD98059 inhibited $TNF-{\alpha}-induced$ LOX-1 expression. CPT and NAC suppressed $TNF-{\alpha}-induced$ LOX-1 expression and phosphorylation of $NF-{\kappa}B$ p65 and ERK in rat aorta. These data suggested that $TNF-{\alpha}$ induced LOX-1 expression via ROS activated $NF-{\kappa}B/ERK$ pathway, which could be inhibited by CPT. This study provides new insights for the anti-atherosclerotic effect of CPT.

Analysis of load-settlement behaviour of shallow foundations in saturated clays based on CPT and DPT tests

  • Mir, Mouna;Bouafia, Ali;Rahmani, Khaled;Aouali, Nawel
    • Geomechanics and Engineering
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    • v.13 no.1
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    • pp.119-139
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    • 2017
  • Static Penetration Test (CPT) and Dynamic Penetration Test (DPT) are commonly used in-situ tests in a routine geotechnical investigation. Besides their use for qualitative investigation (lithology, homogeneity and spatial variability), they are used as practical tools of geotechnical characterization (resistance to the penetration, soil rigidity) and modern foundation design as well. The paper aims at presenting the results of an extensive research work on the evaluation of the 1D primary consolidation settlement of saturated clayey soils on the basis of the CPT or DPT tests. The work is based on an analysis of the correlations between the tip resistance to penetration measured in these tests and the parameters of compressibility measured by the compressibility oedometer test, through a local geotechnical database in the northern Algeria. Such an analysis led to the proposal of two methods of calculation of the settlement, one based on the CPT test and the other one on the DPT. The comparison between the predicted settlements and those computed on the basis of the oedometer test showed a good agreement which demonstrate the possbility to use the CPT and DPT tests as reliable tools of computation of foundation settlements in clayey soils.

A study on data mining techniques for soil classification methods using cone penetration test results

  • Junghee Park;So-Hyun Cho;Jong-Sub Lee;Hyun-Ki Kim
    • Geomechanics and Engineering
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    • v.35 no.1
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    • pp.67-80
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    • 2023
  • Due to the nature of the conjunctive Cone Penetration Test(CPT), which does not verify the actual sample directly, geotechnical engineers commonly classify the underground geomaterials using CPT results with the classification diagrams proposed by various researchers. However, such classification diagrams may fail to reflect local geotechnical characteristics, potentially resulting in misclassification that does not align with the actual stratification in regions with strong local features. To address this, this paper presents an objective method for more accurate local CPT soil classification criteria, which utilizes C4.5 decision tree models trained with the CPT results from the clay-dominant southern coast of Korea and the sand-dominant region in South Carolina, USA. The results and analyses demonstrate that the C4.5 algorithm, in conjunction with oversampling, outlier removal, and pruning methods, can enhance and optimize the decision tree-based CPT soil classification model.

Sequential administration of camptothecin sensitizes human colon cancer HCT116 cells to paclitaxel via $p21^{Cip1/WAF1}$

  • Yoo, Jung-Min;Kim, Yun-Jin;Lee, Sung-Jae;Kim, Sang-Hoon
    • Animal cells and systems
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    • v.15 no.1
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    • pp.9-17
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    • 2011
  • Colorectal cancer is the third leading cause of cancer-related death in Western countries. Chemotherapeutic agents with different mechanisms of action have shown an increase in cure rates. In the present study, we investigated the effect of a combination of low concentration of paclitaxel (taxol, 5 nM) and topoisomerase 1 inhibitor camptothecin (CPT) on HCT116 colon cancer cells. Although the viability of cells treated with taxol alone was similar to that of control cells, sequential treatment with taxol and CPT exhibited high cytotoxicity. However, the opposite sequence of treatment did not exert cytotoxic effects on HCT116 cells. This enhanced cytotoxicity of the sequential combination therapy was the result of mitotic arrest, which increased the level of $p21^{Cip1/WAF1}$ through the p38 mitogen-activated protein kinase (MAPK) pathway. Knockdown by $p21^{Cip1/WAF1}$ siRNA or treatment with a p38 inhibitor reduced the viability of cells sequentially exposed to taxol and CPT. Taken together, a low taxol concentration in combination with CPT induced mitotic arrest in HCT116 cells, leading to synergistic cell death through enhanced expression of $p21^{Cip1/WAF1}$ and p38 MAPK pathway. Therefore, taxol could playa role as a sensitizer of CPT in colon cancer cells.

Fatty acid uptake and oxidation in skeletal muscle

  • Yun, Hea-Yeon;Tamura, Tomohiro;Lim, Kiwon
    • Korean Journal of Exercise Nutrition
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    • v.16 no.1
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    • pp.1-9
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    • 2012
  • Long chain fatty acids (LCFAs) are transported into cells via plasma transporters, are activated to fatty acyl-CoA by fatty acyl-CoA synthase (ACS), and enter mitochondria via the carnitine system (CPT1/CACT/CPT2). The mitochondrial carnitine system plays an obligatory role in β-oxidation of LCFAs by catalyzing their transport into the mitochondrial matrix. Fatty acyl-CoAs are oxidized via the β-oxidation pathway, which results in the production of acetyl-CoA. The acetyl-CoA can be imported into the tricarboxylic acid (TCA) cycle for oxidation in the mitochondrial matrix or can be used for malonyl-CoA synthesis by acetyl-CoA carboxylase 2 (ACC2) in the cytoplasm. In skeletal muscle, ACC2 catalyzes the carboxylation of acetyl-CoA to form malonyl-CoA, which is a potent endogenous inhibitor of carnitine palmitoyltransferase 1 (CPT1). Thus, ACC2 indirectly inhibits the influx of fatty acids into the mitochondria. Fatty acid metabolism can also be regulated by malonyl-CoA-mediated inhibition of CPT1.

RELATIONSHIP BETWEEN CHANGES IN EVENT-RELATED POTENTIALS AND CHANGES IN CONTINUOUS PERFORMANCE TEST UNDER THE INFLUENCE OF METHYLPHENIDATE IN ATTENTION-DEFICIT/HYPERACTIVITY DISORDER (주의력 결핍 ${\cdot}$ 과잉행동장애 아동에서 Methylphenidate에 의한 사건관련전위와 연속과제수행 변화사이의 상관성)

  • Choi, Young;Lee, Mu-Suk;Lee, Mi-Suk
    • Journal of the Korean Academy of Child and Adolescent Psychiatry
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    • v.8 no.2
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    • pp.273-286
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    • 1997
  • Objective:This study was designed to evaluate effects of methylphenidate(MPH) on event-related potentials(ERP) and continuous performance test(CPT) in attention-deficit hyperactivity disorder (ADHD) and to see the correlation between changes in ERP and changes in performance. Method:ERP and CPT were used to examine the acute effects of MPH(0.5mg/kg) in eleven ADHD boys(89-103 months old). Results:1) After MPH administration, P3 latency to nontarget stimuli at Fz was significantly decreased (p<0.01) and P2 amplitudes to target stimuli at Pz and at Oz and P3 amplitude to target stimuli at Cz were significantly increased(p<0.05). 2) Commission error and omission error in the CPT-X and commission error in the CPT-AX were decreased(p<0.01), and hits and perceptual sensitivity(d') in the CPT-X and d′ in the CPT-AX were increased(p<0.01). 3) The change of P3 latency to nontarget stimuli at Fz and the change of d′ in the CPT-X were negatively correlated(p<0.05), and the change of P2 amplitude to target stimuli at Pz and d′ in the CPT-AX were positively correlated(p<0.05). Conclusion:MPH improves change orienting reaction, the delivery of task relevant information, accuracy and perceptual sensitivity in ADHD. And the increase of ability to discriminate targets from non-targets reflects reduced evaluation time in large memory component task and enhanced change orienting reaction in simple task.

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