• Title/Summary/Keyword: leukemia Jurkat T cells

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In Vitro Selection of Cancer-Specific RNA Aptamers

  • Lee Young-Ju;Lee Seong-Wook
    • Journal of Microbiology and Biotechnology
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    • v.16 no.7
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    • pp.1149-1153
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    • 2006
  • In this study, nuclease-resistant RNA aptamers that are specific for Jurkat T leukemia cells were selected by a subtractive systemic evolution of ligands by exponential enrichment (SELEX) method. A randomized nuclease-resistant RNA library was incubated with normal peripheral blood mononuclear cells (PBMC) in each round to preclude RNAs that recognize the common cellular components on the surface of normal and cancer cells. The precluded RNAs were used for the selection of Jurkat T cell-specific aptamers, and the specific RNAs were then gradually enriched from start to the following selections. After 16 rounds of the subtractive SELEX, the selected aptamers were found to preferentially bind to Jurkat T cells, but not to the normal PBMC, evidenced by fluorescence-activated cell sorting analysis. Thus, the subtractive SELEX can be used to identify ligands to cancer-specific biological markers without prior knowledge of the nature of markers. The aptamers could be applied to specific cell sorting, tumor therapy, and diagnosis, and moreover, to find cancer cell-specific markers.

Studies on Production of Monoclonal Antibodies Reactive with T-Cell Leukemia (인형 T세포 백혈병에 대한 단세포군 항체 생산에 관한 연구)

  • 서병석;김원배;최응칠;김병각
    • YAKHAK HOEJI
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    • v.31 no.5
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    • pp.253-265
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    • 1987
  • To develop hybridomas secreting monoclonal antibodies to be used as unlimited sources of reagents indispensable for the diagnosis and treatement of leukemic malignancy, a monoclonal antibody was generated to human pre-T leukemia cells (Jurkat). Hybridomas were produced against Jurkat cell line by fusing spleen cells from hyperimmunized mice with murine plasmacytoma cells (P3$\times$63Ag8. V653). One monoclonal antibody derived from this fusion, designated DMJ-2 was reactive with T-cell lines (Jurkat, Molt-4 and RPMI-8402) and normal peripheral E-rosette forming T cells, but unreactive with B-cell lines (Daudi, Nalm-6) and non-T, non-B cell line (K562). Conclusively DMJ-2 reactive with mature and immature T-lineage lymphoid cells.

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Effect of the Water Extract of Ajbizzia julibrissin on Apoptotic Cell Death in the Human Leukemic Jurkat T Cell Line (백혈병세포주 Jurkat T 세포에서 합환피 (Ajbizzia julibrissin) 물 추출물의 아포토시스 유도 효과)

  • 황상구;이형철;김춘관;김용익;주성민;김원신;전병훈
    • YAKHAK HOEJI
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    • v.45 no.6
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    • pp.730-738
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    • 2001
  • Apoptosis is a morphologically and biochemically distinct form of cell death that occurs in many different cell types in a wide variety of organisms. Ajbizzia julibrissin belonging the family Leguminosae has been used for the treatment of contusion, sore throat, amnesia, and insomnia in oriental traditional medicine. This study investigates whether the water extract off julibrissin induce apoptotic cell death in Jurkat T-acute lymphoblastic leukemia (ALL) cells. Jurkat cells were increased inhibitions of cell viability in a concentration-dependent manner by A julibrissin. This herbal medicine also caused apoptosis as measured by cell morphology and DNA fragmentation. The capability oft julibrissin to induce apoptosis was associated with proteolytic cleavage of specific target protein such as poly (ADP-ribose) polymerase (PARP) protein suggesting the possible involvement of caspases. Our result skewed that Bcl-2 and Bax protein levels were not changed in all A julibrissin-treated groups compared to control group. These results suggest that A julibrissin-mediated apoptosis is independent with Bcl-2 related signaling pathway in this cells.

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Screening of the Extracts of Herbal Medicines which Stimulate the Hydrolysis of Phosphoinositides in Jurkat T-lymphocyte Cells (Jurkat T 면역세포에서 Phosphoinositides의 가수분해를 증가시키는 약용식물 추출물의 검색)

  • 민도식;이영한;백석환;서판길;류성호
    • Biomolecules & Therapeutics
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    • v.4 no.2
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    • pp.148-153
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    • 1996
  • Activation of the T lymphocytes results in a variety of early biochemical events ultimately leading to cell proliferation and lymphokine production. Stimulation of the signal transduction cascade in T cells through the T cell receptor coincides with activation of the phosphatidylinositol-phospholipase C (PI-PLC) pathway. Therefore, we have established a model system to screen immune-simulator that can increase the hydrolysis of phosphoinositides in human T cell leukemia Jurkat cells. As a result of screening from herbal medicine extract, 4 extracts (O1ibanum, Ephedrae Herba, Real Gar, Saussureae Radix) were found 14 increase the production of inositol phosphates. All the active fraction from the four kinds of extract were fluted in a different retention time on C-18 HPLC and these active fraction also showed difference in cell specificity. And all the active fractions increased DNA synthesis in T cell. Therefore, it is suggested that the active fraction among 4 extracts might contain a compound having different properties one another.

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Effect of the Water Extract of Albizzia julibrissin on Cell Cycle Progression in the Human Leukemic Jurkat Cells (백혈병세포주 Jurkat의 세포주기 억제에 미치는 합환피(Albizzia julibrissin) 물 추출물의 효과)

  • Hwang, Sang-Gu;Lee, Hyung-Chul;Kim, Dae-Geun;An, Won-Gun;Jeon, Byung-Hun
    • Korean Journal of Pharmacognosy
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    • v.33 no.1 s.128
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    • pp.29-34
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    • 2002
  • Albizzia julibrissin belonging to the family Leguminosae has been used for the treatment of contusion, sore throat, amnesia, and insomnia in Oriental traditional medicine. The water extract of A. julibrissin induced apoptosis in Jurkat T-acute lymphoblastic leukemia (ALL) cells as measured by cell morphology. The capability of this herb medicine to induce apoptosis was associated with proteolytic cleavage of specific target protein such as beta-catenin protein suggesting the possible involvement of caspases. The purpose of the present study is also to investigate the effect of A. julibrissin on cell cycle progression. Our results showed that GI checkpoint related gene products (cyclin D1, cyclin dependent kinase 4, retinoblastoma, E2F1) were decreased in their protein levels in a dose-dependent manners after treatment of the extract. These results indicate that the increase of apoptotic cell death by A. julibrissin may be due to the inhibition of cell cycle progression in wild type p53-lacking Jurkat cells.

Proteolysis of $\beta$-Catenin in Apoptotic Jurkat Cells

  • Hwang, Sang-Gu;Park, Jeong-Uck;Lee, Hyung-Chul;Joo, Woo-Hong;Cho, Yong-Kweon;Moon, Ja-Young
    • Journal of Life Science
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    • v.10 no.1
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    • pp.57-63
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    • 2000
  • ${\beta}$-catenin, which plays a critical role in both the cytoskeleton and in transcriptional regulation in variousadherent cell types, undergoes degradation during adherent cell apoptosis. Although ${\beta}$-catenin has been reported to be present in Jurkat T-acute lymphoblastic leukemia cells, the regulation of ${\beta}$-catenin in hematologic malignancies have not been examined. The data presented here demonstrate that treatment of the T cell leukemia Jurkat iwht the apoptosis inducer anti-Fas induced proteolytic cleavage of ${\beta}$-catenin. ${\beta}$-catenin was cleaved at both the N- and C-terminus after anti-Fas treatment. Cleavage of intact ${\beta}$-catenin was completely inhibited by caspase selective protease inhibitors. These data demonstrate that ${\beta}$ -catenin proteolysis is triggered by the cross-linking of the Fas receptor on Jurkat cells and subsequent activation of caspase protease. There was a clear accumulatio of the large proteolytic fragment in Jurkat cells treated with lactacystin of ALLM. These are potent inhibitors of proteasome and calpain. these results suggest that both the proteasome and clapain may recognize the large ${\beta}$-catenin fragment as a substrate fot further degradation and that these pathewasy may act downstream of scapase in response to Fas receptor activation. Therefore, we suggest that ${\beta}$-catenin may play a role in promoting Jurkat survival.

Activation of Pro-Apoptotic Multidomain Bcl-2 Family Member Bak and Mitochondria-Dependent Caspase Cascade are Involved in p-Coumaric Acid-Induced Apoptosis in Human Jurkat T Cells (p-Coumaric acid에 의해 유도되는 인체 Jurkat T 세포의 에폽토시스 기전)

  • Lee, Je-Won;Kim, Young-Ho
    • Journal of Life Science
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    • v.21 no.12
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    • pp.1678-1688
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    • 2011
  • The apoptogenic effect of p-coumaric acid, a phenolic acid found in various edible plants, on human acute leukemia Jurkat T cells was investigated. Exposure of Jurkat T cells to p-coumaric acid (50-$150{\mu}M$) caused cytotoxicity and TdT-mediated dUTP nick-end labeling (TUNEL)-positive apoptotic DNA fragmentation along with Bak activation, ${\Delta}{\psi}m$ loss, activation of caspase-9, -3, -7, and -8, and PARP degradation in a dose-dependent manner. However,these apoptotic events were completely abrogated in Jurkat T cells overexpressing Bcl-2.Under these conditions, necrosis was not accompanied. Pretreatment of the cells with the pan-caspase inhibitor (z-VAD-fmk) could prevent p-coumaric acid-induced sub-$G_1$ peak representing apoptotic cells, whereas it failed to block ${\Delta}{\psi}m$ loss, indicating that the activation of caspase cascade was prerequisite for p-coumaric acid-induced apoptosis as a downstream event of ${\Delta}{\psi}m$ loss. FADD- and caspase-8-positive wild-type Jurkat T cell clone A3, FADD-deficient Jurkat T cell clone I2.1, and caspase-8-deficient Jurkat T cell clone I9.2 exhibited similar susceptibilities to the cytotoxicity of p-coumaric acid, excluding an involvement of Fas/FasL system in triggering the apoptosis. The apoptogenic activity of p-coumaric acid is more potent in malignant Jurkat T cells than in normal human peripheral T cells. Together, these results demonstrated that p-coumaric acid-induced apoptogenic activity in Jurkat T cellswas mediated by Bak activation, ${\Delta}{\psi}m$ loss, and subsequent activation of multiple caspases such as caspase-9, -3, -7, and-8, and PARP degradation, which could be regulated by anti-apoptotic protein Bcl-2.

Effect of Ailanthus altissima Water Extract on Cell Cycle Control Genes in Jurkat T Lymphocytes (Jurkat T 임파구의 세포주기 기전에 미치는 저근백피(Ailanthus altissima)의 효과)

  • 전병훈;황상구;이형철;김춘관;김대근;이기옥;윤용갑
    • YAKHAK HOEJI
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    • v.46 no.1
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    • pp.18-23
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    • 2002
  • Ailanthus altissima has been used to settle an upset stomach, to alleviate a fever and as an insecticide. We reported that the water extract of A. altissima induced apoptotic cell death in Jurkat T-acute Iymphoblastic leukemia cells. Here, we showed the dose-dependent inhibitions of cell viability by the extract, as measured by cell morphology. The cell cycle control genes are considered to play important roles in tumorigenesis. The purpose of the present study is also to investigate the effect of A. altissima on cell cycle progression and its molecular mechanism in the cells. The level of p21 protein was increased after treatment of the extract, whereas both Bcl-2 and Bax protein levels were not changed. These results suggest that A. altissima induces apoptotic cell death via p21-dependent signaling pathway in Jurkat cells which delete wild type p53. Gl checkpoint related gene products tested (cyclin D3, cyclin dependent kinase 4, retinoblastoma, E2Fl) were decreased in their protein levels in a dose-dependent manner after treatment of the extract Taken together, these results indicate that the increase of apoptotic cell death by A. altissima may be due to the inhibition of cell cycle in Jurkat cells.

Spatholobus suberectus Water Extract induces Apoptotic Cell Death via Inhibition of Cell Cycle in Jurkat Human Leukemia Cell Line (계혈등 추출물이 Jurkat T 임파구의 세포고사 및 세포주기 억제에 미치는 효과)

  • Cho Nam Su;Jung Woo Cheol;Na Heon Sik;Song Young Jun;Lee Kye Seung;Lee In;Jeon Byung Hun;Moon Byung Soon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.1
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    • pp.101-109
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    • 2004
  • Spatholobus suberectus belonging the family Leguminosae has been used for promoting blood circulation, removing blood stasis, tonifying the blood, relaxing tendons, stopping internal bleeding and eliminating dampness in oriental traditional medicine. This study investigates whether the water extracts of S. suberectus induce apoptotic cell death in Jurkat T-acute lymphoblastic leukemia (ALL) cells. Jurkat cells were increased inhibitions of cell viability in a concentration-dependent manner by S. suberectus, as measured by cell morphology. The capability of S. suberectus to induce apoptosis was associated with proteolytic cleavage of specific target protein such as poly (ADP­ribose)polymerase protein suggesting the possible involvement of caspases. The purpose of the present study is also to investigate the effect of S. suberectus on cell cycle progression. G1 checkpoin related gene products tested (cyclin D1, cyclin dependent kinase 4, retinoblastoma, E2Fl) were decreased in their protein levels in a dose-dependent manners after treatment of the extract. These results indicate that the increase of apoptotic cell death by S. suberectus may be due to the inhibition of cell cycle progression in wild type p53-lacking Jurkat cells.

Machilus Thunbergii Water Extract Induces Cytotoxic Effect against Human Acute Jurkat T Lymphoma (후박 열수 추출물의 Jurkat T 세포에서 세포사멸 효과)

  • Kim, Min Hwan;Lee, Jong-Hwan
    • Journal of Life Science
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    • v.27 no.8
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    • pp.951-957
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    • 2017
  • To understand the cytotoxic activity of Machilus thunbergii, which has been used as a traditional oriental medicine, the mechanism underlying the cytotoxic effect of its extract on human acute Jurkat T cells was investigated. The methanol extract of roots (3 kg) of M. thunbergii was evaporated, dissolved in, and then extracted by water. The water-extracted active substance was designated MTWE. When Jurkat T cells were treated with MTWE at concentrations of 0, 25, 50, and $100{\mu}g/ml$, the apoptotic phenomenon of cells accompanying several subsequent biochemical reactions, such as mitochondrial cytochrome c release, activation of caspase-3, and ICAD degradation, was detected in the Jurkat T cells. Moverover. the expression of Bcl-xL, which is a suppressor for mitochondrial cytochrome c release pathway, was reduced in the Jurkat T cells. As DUSP6, a growth suppressor of cancer cells, ranged from 0, 25, 50, $100{\mu}g/ml$ of MTWE, the expression level was elevated in the Jurkat T cells. The apoptotic morphological change of the nuclei was observed by DAPI staining. Although the potential involvement of the other factors and DUSP6 is currently being investigated in more detail, these findings support the notion that MTWE is able to achieve the apoptosis of Jurkat T cells, and it seems that MTWE is useful as a method of evaluating a chemotherapeutic agent or tonic materials for human acute leukemia.