• 한국생물정보학회:학술대회논문집
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• 한국생물정보시스템생물학회 2001년도 제2회 생물정보 워크샵 (DNA Chip Bioinformatics)
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• pp.29-44
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• 2001

DNA damage by physical insult including UV and g-radiation might provoke genetic alterations in cells, which is followed by either acute cell death or tumorigenesis. The responsiveness to g-radiation depends on cellular context of target cells. To understand the mechanisms of checkpoint control, repair and cell death following genotoxic stimu]i, cDNA microarray can provide the gene expression profile. To make a profile of gene expression in irradiated Jurkat T cells, we hybridized the cDNA microarray using cDNA from g-irradiated Jurkat T cells. Jurkat T cells were exposed to 4Gy to 16Gy, and total RNA were extracted at 4 to 24 hrs after irradiation. The hybridization of the microarray to fluorescence-labeled cDNA from treated and untreated cells was analyzed by bioinformatic analysis to address relative changes in expression levels of the genes present in the array. Responses varied widely in different time points, suggesting acute stress response and chronic restoration or cell death. From these results we could select 384 genes related to radiation response in Tcells, and radiation response might be different in various types of cells. Using Radchip, we could separate "the exposed" from control PBMCs. We propose that Radchip might be useful to check the radiation research as well as radiation carcinogenesis.

• 한국균학회지
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• 제27권4호통권91호
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• pp.312-317
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• 1999

우리나라 중요한 식품 원료인 전통 매주로부터 분리한 단독균의 접종 메주의 암세포 저해효과를 검색하기 위하여, 민간유래의 혈액암 세포주에 대한 저해활성을 MTT assay로 분석하였다. 먼저 전통 메주로부터 21종의 단독균을 분리한 후 각각 접종하여 발효된 단독균의 메주시료를 조제한 다음 80% methanol로 추출하였다. 메주 메탄올추출물은 혈액암세포중 HL60에서는 다소 낮은 성장 저해효과를 보였으나, U937과 Jurkat cell에서는 저해효과가 큰 것으로 나타났다. 특히 Mucor속과 Absidia속 Aspergillus속으로 제조된 메주들에서 이들 혈액암세포에 대해 저해효과가 큰 것으로 나타났다. 그러나 모든 메주 메탄올추출물들은 인간의 정상 lymphocyte에서 대해서는 90% 이상의 높은 생존율을 나타내어 정상 세포에 대한 성장 저해 효과가 거의 없음을 보며주었다. 이는 단독균의 메주시료가 가지는 세포독성이 암세포에 대한 특이적인 작용인 것으로 나타났다.

• 생명과학회지
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• 제19권11호
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• pp.1529-1537
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• 2009

L-arginine 구조유사체인 L-canavanine의 인체 급성백혈병 Jurkat T 세포에 대한 apoptosis 유도활성이 단백질 티로신키나아제 $p56^{lck}$에 어떻게 조절되는지를 규명하기 위해 $p56^{lck}$를 발현하는 Jurkat T 세포주 E6.1과 $p56^{lck}$-결손 Jurkat T 세포주 JCaM1.6에 있어서 L-canavanine의 세포독성, L-canavanine에 의해 유도되는 apoptotic DNA fragmentation 및 apoptotic sub-$G_1$ peak를 비교하여 본 바, $p56^{lck}$-negative JCaM1.6 세포가 $p56^{lck}$-positive E6.1 세포에 비해 L-canavanine의 apoptotis 유도활성에 훨씬 더 민감한 것으로 나타났다. 이러한 $p56^{lck}$-negative JCaM1.6 세포의 민감성은 JCaM1.6 세포에 $p56^{lck}$ 유전자를 transfection시켜 발현시키면 현저히 감소되었다. L-Canavanine에 의해 유도되는 apoptosis관련 현상들을 $p56^{lck}$-stable transfectant인 JCaM1.6/lck 세포와 empty vector-transfectant 인 $p56^{lck}$-negaive JCaM1.6/vector 세포에서 Western blot analysis로 비교한 결과, L-canavanine에 의해 유도되는 mitochondrial membrane potential (${\Delta\Psi}m$)의 감소, caspase-9, -8, -7 및 -3의 활성화, 그리고 PARP 및 $PLC{\gamma}$-1의 분해가 JCaM1.6/vector 세포에 비해 JCaM1.6/lck 세포에서 더 약하게 나타났다. JCaM1.6/lck 세포를 2.5 mM L-canavanine으로 처리한 다음 세포 내 $p56^{lck}$ kinase 활성의 변화를 $\alpha$-casein을 기질로 하여 시간 별로 측정한 결과, L-canavanine의 처리 후 15분만에 $p56^{lck}$ kinase의 활성이 약 1.6배 증가되었으며 이후 6시간 동안은 약 1.3~1.4 배정도 증가된 수준으로 kinase 활성이 유지되는 것으로 확인되었다. L-Canavanine에 의한 apoptosis의 개시에 Fas/FasL 상호작용이 관련되는지를 규명하기 위해 FADD-negative Jurkat T 세포주 I2.1, caspase-8-negative Jurkat T 세포주 I9.2 및 wild-type Jurkat T 세포주 A3에 대한 L-canavanine의 세포독성을 비교한 결과, A3와 I2.1 세포의 경우는 L-canavanine의 세포독성이 동일하게 나타났고, 특히 caspase-8가 결손된 I9.2 세포의 경우는 L-canavanine의 세포독성에 대한 민감성이 A3와 I2.1 세포에 비해 단지 미약하게만 완화되는 것으로 나타나, L-canavanine의한 apoptosis에는 Fas/FasL 상호작용이 관련되어 있지 않으며, 또한 caspase-8의 역할이 필수적이지 않음을 시사하였다. Jurkat T 세포에 있어서 L-canavanie에 의해 유도되는 sub-$G_1$ peak 및 caspases 활성화에 미치는 pan-caspase inhibitor (z-VAD-fmk), caspase-9 inhibitor (z-LEHD-fmk), caspase-3 inhibitor (z-DEVD-fmk), caspase-4 inhibitor (z-LEVD-fmk) 및 caspase-12 inhibitor (z-ATAD-fmk)의 영향을 조사한 결과, L-canavanie에 의한 apoptosis는 ${\Delta\Psi}m$의 감소, caspase-9 및 caspase -3의 활성화에 뒤따른 caspase-8 및 caspase-7의 활성화, 그리고 PARP의 분해의 순서로 유도되는 것으로 나타났으며, 아울러 caspase-9의 활성화와 함께 caspase-12의 활성화가 L-canavanine 처리에 따른 caspase-3의 활성화에 요구되는 것으로 확인되었다. 결론적으로, L-canavanine 처리에 의한 Jurkat T 세포의 apoptosis는 ${\Delta\Psi}m$ 감소, caspase-9, caspase-3 및 caspase-7의 활성화에 의해 유도되며, 이들 apoptosis 현상들은 $p56^{lck}$에 의해 negative regulation되었다.

• Journal of Periodontal and Implant Science
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• 제27권4호
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• pp.805-818
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• 1997

Immune responses of periodontal tissue may be regulated by products of sensory afferent nerve endings such as neuropeptides. Substance P(SP), a tachykinin neuropeptide, has been previously reported to stimulate the activities of T lymphocyte. Therefore, I examined the role of SP in IL-2 production and cell proliferation by using a homogeneous line of T lymphocytes(Jurkat and HuT78). Cell proliferation rate was determined by [$^3H$]-thymidine incorporation test, and IL-2 was quantitated by the growth rate of CD4+ IL-2-dependent T lymphocyte line CTLL-2. SP stimulated cell proliferation of T lymphocytes at the concentration of $10^{-12}$ and $10^{-8}$M in a biphasic bell-shape dose-dependent manner. However, SP alone did not induce IL-2 release at the concentration range of $10^{-6}$ to $10^{-14}$M. The upregulation of IL-2 release was observed when $10^{-12}$M SP was applied together with mitogens such as Con A or PHA+PMA on T cell lines, especially on Jurkat. Con A or PHA+PMA demonstrated to increase the rate of cell proliferation of Jurkat, which had shown to produce much amount of IL-2 indicating that mitogen-induced cell proliferation might be partially influenced by released IL-2. It was concluded that regulatory effects of SP on the immune/inflammatory response could be mediated through the costimulatory upregulation of IL-2 production and increase of cell proliferation of T lymphocyte.

• Journal of Microbiology and Biotechnology
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• 제30권12호
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• pp.1885-1895
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• 2020

Rumex japonicus Houtt (RJH) is a valuable plant used in traditional medicine to treat several diseases, such as scabies and jaundice. In this study, Jurkat cell growth inhibitory extracts of R. japonicus roots were subjected to bioassay-guided fractionation, resulting in the isolation of three naphthalene derivatives (3-5) along with one anthraquinone (6) and two phenolic compounds (1 and 2). Among these compounds, 2-methoxystypandrone (5) exhibited potent anti-proliferative effects on Jurkat cells. Analysis by flow cytometry confirmed that 2-methoxystypandrone (5) could significantly reduce mitochondrial membrane potential and promote increased levels of mitochondrial reactive oxygen species (ROS), suggesting a strong mitochondrial depolarization effect. Real-time quantitative polymerase chain reaction (qPCR) analysis was also performed, and the results revealed that the accumulation of ROS was caused by reduced mRNA expression levels of heme oxygenase (HO-1), catalase (CAT), glutathione peroxidase (GPx), and superoxide dismutase (SOD). In addition, 2-methoxystypandrone (5) triggered strong apoptosis that was mediated by the arrest of the G0/G1 phase of the cell cycle. Furthermore, 2-methoxystypandrone (5) downregulated p-IκB-α, p-NF-κB p65, Bcl2, and Bcl-xl and upregulated BAX proteins. Taken together, these findings revealed that 2-methoxystypandrone (5) isolated from RJH could potentially serve as an early lead compound for leukemia treatment involving intracellular signaling by increasing mitochondrial ROS and exerting anti-proliferative effects.

• Journal of Microbiology and Biotechnology
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• 제10권1호
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• pp.27-34
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• 2000

The chloroform and methanol (2;1, v/v) extract from an edible plant, Actinidia arguta Planchon, appeared to possess antitumor activity against human leukemias Jurkat T and U937 cells through inducing apoptosis. The substance in the solvent extract was purified by silica gel column chromatography, preparative TLC, and Sephadex LH-20 column chromatography. Characteristics of the substance analyzed by UV scanning analysis, $^1H$ and $^{13}C$ NMR spectra suggested that the substance belongs to the chlorophyll derivatives-like group. The $IC_{50}$ value of the chlorophyll derivative (Cp-D) determined by MTT assay was $15\mu\textrm{g}/ml$ for Jurkat, $10\mu\textrm{g}/ml$ for U937, and $11.4\mu\textrm{g}/ml$ for HL-60m and was more toxic to these leukemias than to solid tumors or normal fibroblast. In order to elucidate cellular mechanisms underlying the cytotoxicity, the effect of the Cp-D on Jurkat T cells was investigated. When cells were treated with the Cp-D at a concentration of $15\mu\textrm{g}/ml$, [3H]thymidine incorporation declined rapidly and wa undetectable in 1h. However, no significant changes were made in the cell cycle distribution of the cells by 24h. The sub-Gl peak representing apoptotic cells began to be detectable in 36h, at which time apoptotic DNA fragmentation was also detected on agarose gel electrophoresis, demonstrating that the cytotoxic effect of the Cp-D is attributable to the induced apoptosis. Under the same conditions, although the protein level of cyclin-dependent kinases such as cdc4, csk6, cdk2, and cdc2 was not significantly changed until 24h, the kinase activity of all c안 rapidly declined and reached a minimum level within 1-6h and then recovered to the initial level by 12h and sustained until 24h. These results suggest that inactivation of cdks at an inappropriate time during the cell cycle progression in jurkat T cells following a treatment with the Cp-D leads to induction of apoptotic cell death.

• Journal of Microbiology and Biotechnology
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• 제27권1호
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• pp.197-205
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• 2017

Exposure of Jurkat T cell clone (J/Neo cells) to acacetin (5,7-dihydroxy-4'-methoxyflavone), which is present in barnyard millet (Echinochloa esculenta (A. Braun)) grains, caused cytotoxicity, enhancement of apoptotic $sub-G_1$ rate, Bak activation, loss of mitochondrial membrane potential (${\Delta}{\Psi}m$), activation of caspase-9 and caspase-3, degradation of poly(ADP-ribose) polymerase, and FITC-Annexin V-stainable phosphatidylserine exposure on the external surface of the cytoplasmic membrane without accompanying necrosis. These apoptotic responses were abrogated in Jurkat T cell clone (J/Bcl-xL) overexpressing Bcl-xL. Under the same conditions, cellular autophagic responses, including suppression of the Akt-mTOR pathway and p62/SQSTM1 down-regulation, were commonly detected in J/Neo and J/Bcl-xL cells; however, formation of acridine orange-stainable acidic vascular organelles, LC3-I/II conversion, and Beclin-1 phosphorylation (Ser-15) were detected only in J/Neo cells. Correspondingly, concomitant treatment with the autophagy inhibitor (3-methyladenine or LY294002) appeared to enhance acacetin-induced apoptotic responses, such as Bak activation, ${\Delta}{\Psi}m$ loss, activation of caspase-9 and caspase-3, and apoptotic $sub-G_1$ accumulation. This indicated that acacetin could induce apoptosis and cytoprotective autophagy in Jurkat T cells simultaneously. Together, these results demonstrate that acacetin induces not only apoptotic cell death via activation of Bak, loss of ${\Delta}{\Psi}m$, and activation of the mitochondrial caspase cascade, but also cytoprotective autophagy resulting from suppression of the Akt-mTOR pathway. Furthermore, pharmacologic inhibition of the autophagy pathway augments the activation of Bak and resultant mitochondrial damage-mediated apoptosis in Jurkat T cells.

• 생명과학회지
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• 제19권2호
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• pp.163-168
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• 2009

천초근은 전통적으로 동양의학에서 항암제로 사용되어왔는데 인간 급성 백혈병 세포주인 Jurkat T 세포를 사용하여 천초근의 세포독성기작을 알아보았다. 천초근 뿌리(3 kg)를 메탄올로 추출, 증류한 후 물에 녹여 다시 dichloromethane으로 추출분획 하였다. 세포독성 활성을 보이는 dichloromethane 추출물을 연속적으로 HPLC를 통해 분리하였고 그 활성물질(65 mg)을 CCH1이라 명명하였다. CCH1을 0.5 ${\mu}g$/ml에서 2.0 ${\mu}g$/ml의 농도로 처리하고 세포사멸 과정을 보았다. 즉, mitochondria cytochrome c 방출, casapase-8, -9 및 caspase-3의 활성화, PARP 분해, DNA 단편화 현상들이 일어나는 것을 관찰하였다. 하지만, mitochondria cytochrome c 방출 억제자인 Bcl-xL이 과발현되는 Jurkat T 세포에서는 세포사멸현상이 일어나지 않았다. 이러한 결과는 CCH1이 mitochondria 의존적인 신호전달 과정을 통해서 세포사멸을 유도 한다고 할 수 있다. 그리고 CCH1에 의한 세포독성은 혈액에서 분리한 단핵구 세포보다 Jurkat T 세포에서 보다 강한 활성을 보였다.

• 대한의생명과학회지
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• 제25권1호
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• pp.66-74
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• 2019

Hyperlipidemia is defined as conditions of the accumulation of lipids such as free fatty acids (FFA), triglyceride (TG), cholesterol and/or phospholipid in the bloodstream. Hyperlipidemia can cause lipid accumulation in non-adipose tissue, which is lipid-cytotoxic effects in many tissues and mediates cell dysfunction, inflammation or programmed cell death (PCD). TG is considered to be a major cause of atherosclerosis through inflammatory necrosis of vascular endothelial cells. Recently, TG have also been shown to exhibit lipid-cytotoxicity and induce PCD. Therefore, we investigated the effect of TG on the cytotoxic effect of various cell types. When exposed to TG, the cell viability of U937 monocytes and Jurkat T lymphocytes, as well as the cell viability of MCF-7, a non-immune cell, decreased in time- and dose-dependent manner. In U937 cells and Jurkat cells, caspase-9, an intrinsic apoptotic caspase, and caspase-8, an extrinsic apoptotic caspase, were increased by exposure to TG. However, in TG-treated MCF-7 cells, caspase-8 activity increased only without caspase-9 activity. In addition, the reduction of cell viability by TG was recovered when all three cell lines were treated with pan-caspase inhibitor. These results suggest that activation of apoptotic caspases by TG causes lipotoxic effect and decreases cell viability.

• Bulletin of the Korean Chemical Society
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• 제35권1호
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• pp.129-134
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• 2014

Two series of SAHA-liked hydroxamate analogues were designed, synthesized and evaluated for their biological activities against nuclear HDACs. Compounds of Series I were found to be very effective inhibitors of cancer cell growth in the PC-3, Hut78, K562 and Jurkat E6-1 cancer cell lines with mean $IC_{50}$ values from $0.54{\mu}M$ (Ic, Jurkat E6-1) to $7.73{\mu}M$ (Ib, K562), indicating that they are cell permeable and the benzimidazolyl-based ligands are flexible enough to occupy the binding site of HDAC.