• Title/Summary/Keyword: T24 cell

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Induction of apoptosis by water extract Glycyrrhizae radix in human bladder T24 cancer cells (인체 방광암 T24 세포에서 Glycyrrhizae radix 열수추출물에 의한 apoptosis 유도)

  • Eom, Jung Hye;Hwang, Buyng Su;Jeong, Yong Tae;Kim, Min-Jin;Shin, Su Young;Kim, Chul Hwan;Lee, Seung Young;Choi, Kyung Min;Cho, Pyo Yun;Jeong, Jin-Woo;Oh, Young Taek
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.04a
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    • pp.111-111
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    • 2019
  • Glycyrrhizae radix is one of the most frequently prescribed ingredients in Oriental medicine, and G. radix extract has been shown to exert anti-cancer effects. However, the cellular and molecular mechanisms of apoptosis by G. radix are poorly defined. In the present study, it was examined the biochemical mechanisms of apoptosis by water extract of G. radix (WEGR) in human bladder T24 cancer cells. It was found that WEGR could inhibit the cell growth of T24 cells in a dose-dependent manner, which was associated with the induction of apoptotic cell death, as evidenced by the formation of apoptotic bodies, DNA fragmentation and increased populations of annexin-V positive cells. The induction of apoptotic cell death by WEGR was connected with an up-regulation of pro-apoptotic Bax protein expression and down-regulation of anti-apoptotic Bcl-2 and Bcl-xL proteins, and inhibition of apoptosis family proteins (XIAP, cIAP-1 and cIAP-2). In addition, apoptosis-inducing concentrations of WEGR induced the activation of caspase-9, an initiator caspase of the mitochondrial-mediated intrinsic pathway, and caspase-3, accompanied by proteolytic degradation of poly (ADP-ribose)-polymerase. WEGR also induced apoptosis via a death receptor-mediated extrinsic pathway by caspase-8 activation, resulting in the down-regulation of total Bid and suggesting the existence of cross-talk between the extrinsic and intrinsic pathways. Taken together, the present results suggest that WEGR may be a potential chemotherapeutic agent for the control of human bladder cancer cells.

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MST1R as a potential new target antigen of chimeric antigen receptor T cells to treat solid tumors

  • Wen An;Ju-Seop Kang;Sukjoong Oh;Ang Tu
    • The Korean Journal of Physiology and Pharmacology
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    • v.27 no.3
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    • pp.241-256
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    • 2023
  • Although chimeric antigen receptor T cell (CAR-T) is a promising immunotherapy in hematological malignancies, there remain many obstacles to CART cell therapy for solid tumors. Identifying appropriate tumor-associated antigens (TAAs) is especially critical for success. Using a bioinformatics approach, we identified common potential TAAs for CAR-T cell immunotherapy in solid tumors. We used the GEO database as a training dataset to find differentially expressed genes (DEGs) and verified candidates using the TCGA database, obtaining seven common DEGs (HM13, SDC1, MST1R, HMMR, MIF, CD24, and PDIA4). Then, we used MERAV to analyze the expression of six genes in normal tissues to determine the ideal target genes. Finally, we analyzed tumor microenvironment factors. The results of major microenvironment factor analyses showed that MDSCs, CXCL1, CXCL12, CXCL5, CCL2, CCL5, TGF- β, CTLA-4, and IFN-γ were significantly overexpressed in breast cancer. The expression of MST1R was positively correlated with TGF- β, CTLA-4, and IFN-γ. In lung adenocarcinoma, MDSCs, Tregs, CXCL12, CXCL5, CCL2, PD-L1, CTLA-4, and IFN-γ were significantly overexpressed in tumor tissues. The expression of MST1R was positively correlated with TGF- β, CTLA-4, and IFN-γ. In bladder cancer, CXCL12, CCL2, and CXCL5 were significantly overexpressed in tumor tissues. MST1R expression was positively correlated with TGF- β. Our results demonstrate that MST1R has the potential as a new target antigen for treating breast cancer, lung adenocarcinoma, and bladder cancer and may be used as a progression indicator for bladder cancer.

Identification of Biomarkers for Radiation Response Using cDNA Microarray

  • Park, Woong-Yang
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2001.10a
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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.

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The effects of sodium fluoride on oral normal cell cultured in vitro (체외에서 배양된 구강 내 정상세포에 불화나트륨이 미치는 영향)

  • Choi, Byul-Bora;Kim, Da-Hye;Kim, Ji-Young;Park, Sang-Rye
    • Journal of Korean society of Dental Hygiene
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    • v.16 no.3
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    • pp.471-477
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    • 2016
  • Objectives: Fluoride is widely used in the prevention and control of dental caries. The purpose of this study is to examine the biological effects of Sodium fluoride on the proliferation of oral normal cell in vitro(MDPC-23, HaCaT, HGF-1 cells). Methods: The proliferation of normal cells and the cyto-skeletal change of normal cells were assessed by WST-1 assay and F-actin stain assay. The statistical significances of the resulting data were analyzed using SPSS(Window 12.0). Results: The sodium fluoride(0-12 mM) treatment decreased the cell viability in a dose and time dependent manner: HaCaT(6 h): $100{\pm}0$, $98{\pm}0.39$, $82{\pm}2.68$, $75{\pm}0.83$, $69{\pm}1$, $67{\pm}1.42%$(p<0.005); HaCaT(24 h): $100{\pm}0$, $98{\pm}1.85$, $54{\pm}0.64$, $43{\pm}0.4$, $38{\pm}0.32$, $36{\pm}0.13%$(p<0.006), MDPC-23(6 h): $100{\pm}0$, $93{\pm}1.48$, $85{\pm}0.28$, $82{\pm}1.58$, $79{\pm}1.48$, $76{\pm}1.93%$(p<0.009); MDPC-23(24 h): $100{\pm}0$, $91{\pm}1.26$, $58{\pm}0.65$, $49{\pm}1$, $44{\pm}0.74$, $2{\pm}0.05%$(p<0.005), HGF-1(6 h): $100{\pm}0$, $97{\pm}2.93$, $89{\pm}5$, $71{\pm}5.42$, $58{\pm}4.82$, $43{\pm}3.47%$(p<0.009); HGF-1(24 h): $100{\pm}0$, $97{\pm}2.05$, $73{\pm}1.73$, $22{\pm}1.61$, $14{\pm}1.73$, $7{\pm}0.85%$(p<0.005). Thus, changes in cell morphology and disruption of filamentous(F)-actin organization were observed in higher concentration. Conclusions: These results suggest that higher concentrations of fluoride lead to a reduce the number of cells and morphology change of normal cell.

Influence of Water Temperature on Spawning Induction, Egg Development and Seed Collection of Sea Squirt, Halocynthia roretzi (우렁쉥이, Halocynthia roretzi의 산란유발, 난발생 및 채묘에 미치는 수온의 영향)

  • Yoo Sung Kyoo;Kang Kyoung Ho;Chang Young Jin
    • Journal of Aquaculture
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    • v.3 no.1
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    • pp.79-88
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    • 1990
  • In order to obtain the basic data fir the effective seed production of sea squirt, Halocynthia roretzi(Drasche), the influence of water temperature and light intensity on spawning induction, egg development and seed settling were investigated during the Period from December 24, 1906 to February 24, 1987. The maximum number of eggs spawned was obtained at $14^{\circ}C$. The relationships between the water temperature (T, $^{\circ}C$) and the required time (h, hour) in each egg developmental stage were given as follows : Up to 8 cell 1/h =0.0147 T-0.0069(.=0.9816), Up to 32 cell 1/h =0.0100 T-0.0017(r=0.9672), Up to early tadpole 1/h =0.0043 T+0.0024(r=0.9913), Up to hatching larva 1/h =0.0021 T-0.0021 (r=0.9898). The highest rate of seed attachment was obtained at $14^{\circ}C$ of water temperature.

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Insulin - Like Growth Factor-I Effects on the Proliferation and Bone Matrix Protein Gene Expression of MC3T3-E1 Cell (MC3T3-E1 세포증식 및 골기질 단백질 발현에 대한 인슐린유사성장인자-I의 효과)

  • Lee, Dong-Sik;Lee, Jae-Mok;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.30 no.2
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    • pp.389-405
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    • 2000
  • The purpose of this study is to evaluate the effect of IGF-I for DNA synthetic activity and the mRNA expression of bone matrix protein, type I collagen and osteopontin in prolifetation and differentiation of MC3T3-E1 cells. To evaluate DNA synthetic activity, cells were seeded at $2{\times}10^4cells/ml$ in 24 well plates and to evaluate mRNA of type I collagen and osteopontin cells were seeded at $5{\times}10^5cells/ml$ in 100mm culture dishes. These cells were cultured in alpha-minimum essential medium(${\alpha}-MEM$) containing 10% fetal bovine serum at $37^{\circ}C$, 5% $CO_2$ incubator. For DNA synthetic activity test 1, 10, 100ng/ml IGF-I were added to the cells which had been cultured for 3 days before 24 hours. For type I collagen mRNA expression 1, 10ng/ml IGF-I were added to the cells which had been cultured for 5, 10 days and for osteopontin mRNA expression 0.1, 1, 10ng/ml IGF-I were added to the cells which had been cultured for 5, 15, 20 days. Cell proliferaton was measured by the incorporation of [$^3H$]-thymidine into DNA and expression for type I collagen and osteopontin were measured by northern blot analysis. The results were as follows : DNA synthetic activity were generally higher in experimental group than control group. Expressions of type I collagen mRNA were higher at 5 day group and much lower at 10 day group in the control groups. In the experimental groups, mRNA expressions were slightly increased when 1 ng/ml IGF-I were added to 5 day group and decreased in all experimental 10 day groups. Expressions of osteopontin mRNA were higher at 20 day groups and lower at 15 day groups than the control groups. In the experimental groups, mRNA expressions were incereased when 0.1, 1 ng/ml IGF-I were added to 5 day group and in all the 15 day groups, but decreased when 0.1, 1, 10 ng/ml IGF-I were added to 20 day groups. IGF-I stimulated DNA synthetic activity of MC3T3-E1 cells during proliferation stage significantly, did not greatly changed effects on type I collagen mRNA expression and stimulated osteopontin mRNA expression at 15 day especially. In conclusion, we suggests that IGF-I have a tendency of stimulation effect of DNA synthetic activity but do not stimulate type I collagen mRNA in proliferation stage of MC3T3-E1 cell cultures, and stimulate osteopontin mRNA in differentiation stage of MC3T3-E1 cell cultures.

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The Production of Tissue Type-Plasminigen Activator and Mechanism of Cell Death from Human Promyelocytes(HL-60) in Low Serum Containing Medium (저혈청 배지에서 인간 전골수세포(HL-60)를 이용한 tPA 생산과 세포사멸기작에 관한 연구)

  • Kim, Hyun-Gu;Sung, Ki-Don;Kim, Tae-Ho;Ahn, Ju-Hee;Ham, Moon-Sun;Park, Jin-Seo;Lee, Hyeon-Yong
    • Microbiology and Biotechnology Letters
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    • v.24 no.1
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    • pp.126-131
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    • 1996
  • HL-60 was cultivated to produce tPA (tissue-type plasminogen activator) and study the mechanism of cell death. Maximum cell density and tPA production were obtained as $5.27{\times}10^6$ cells/ml and 324ng/ml, respectively under perfusion cultivation. tPA production was enhanced to 420ng/ml in adding 160nM of phorbol ester. The cells were gradually differentiated to granulocytes rather than proliferation. By Fluorescent microscope, apoptosis was prevailed except the death phase and in high agitation speed, but necrosis was prevailed in thawed cells and during the latter periods of the cultivation. It was also proved that tPA was most produced in apoptosis. To obtain higher tPA productivity, the cells must be maintained in apoptosis, not necrosis phase when the cells were dying.

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The Cytotoxic Effects of Paraquat and Bentazon Compensatory Effects of 3-Methylcholanthrene on Kindney of the Rat (제초제 Paraquat와 Bentazon의 세포독성과 3-Methylcholanthrene의 독성경감효과)

  • 임요섭;서대호;한두석
    • Toxicological Research
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    • v.17 no.2
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    • pp.123-129
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    • 2001
  • This study were carried out to investigate cytotoxicity of paraquat and bentazon that is scattering to farm products were essensial for human diet and compensatory effects of 3-methylcholanthrene (3-MC) in vitro and in vivo. In vitro, The 5.0$\times$$10^4$ cell/ml of NIH 3T3 fibroblast in each well of 24 multidish were cultured. After 24 hours, the cells were treated with solution of paraquat and bentazon (1, 25, 50, 100 pM respectively). After the NIH 3T3 fibroblast of all groups were cultured in same condition for 48 hours, Sulfohordamin B Protein (SRB) assay were performed to evaluate the cytotoxicity of cell organelles. Paraquat and bentazon $SRB_50$ were 1860.73 $\mu\textrm{M}$, 1913.38 $\mu$M respectively. In vivo, Sprague Dawley male rats divided into paraquat and bentazon only administered group and simultaneous application group of paraquat and bentazon and 3-MC. At 30 min. and 1, 3, 6, 12, 24, 48 and 96 hrs. interval after each treatment, the animals were sacrificed by decapitation and kidney were immediately removed, immersed in fixatives, and processed with routine method for light microscopic study. Paraffin sections were stained with H-E, PAM, and PAS. Under the light microscope, atrophic change of renal corpuscles were frequently observed from 3 hrs after paraquat and bentazon treatment. The increase of the mesangium was apparent from 12 hrs later after paraquat and bentazon treatment. Necrotic changes of the epithelium and loss of brush border of proximal tubules were most severe at 48 hrs after paraquat and bentazon treatment, respectively. In contrast there were no evidences of the toxic effects on renal tissues at 48hrs in paraquat and bentazon plus 3-MC treated groups.

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Effects of Rubus coreanus and Artemisia princeps Extracts on the Ultraviolet B-Induced DNA Damage Responses in HaCaT Cells (HaCaT 세포에서 자외선 B에 의해 유도된 DNA 상해반응에 대한 복분자와 쑥 추출물의 효과)

  • Lee, Seok Hee;Ha, Se Eun;Lee, Jun Kyoung;Park, Jong Kun
    • Journal of Life Science
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    • v.24 no.2
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    • pp.112-117
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    • 2014
  • We investigated the effects of extracts from Rubus coreanus (RC) and Artemisia princeps var. orientalis (AP) on DNA damage response in ultraviolet B (UVB)-exposed HaCaT cells. Cell activity upon treatment for 24 h with RC or AP alone was similar to or greater than that of the nontreated control. When UVB-exposed cells were postincubated for 24 h in medium containing RC or AP, cell activity increased in a concentration-dependent manner. Nuclear fragmentation analysis showed that postincubation with RC or AP decreased UVB-induced apoptosis by about 20% and 15%, respectively, of that in cells postincubated with growth medium. When UVB-exposed cells were postincubated for 24 h in medium containing RC or AP, the level of cyclobutane pyrimidine dimer decreased in a concentration- dependent manner. Western blot analysis showed that treatment of cells not exposed to UVB with RC or AP alone did not significantly change the levels of phospho-p53 and GADD45 protein. Interestingly, when UVB-exposed cells were postincubated for 24 h in medium containing RC or AP, phospho-p53 and GADD45 levels decreased in a concentration dependent manner. Our results suggest that RC and AP extract assist the survival of UVB-exposed cells in parallel with a decrease in levels of UVB-induced DNA damage and damage-response proteins, such as p53 and GADD45.

Interaction of HIV-1 Core p24 Antigen with Human Monocytic Cell Line THP1 Results in TNF-${\alpha}$ Dependent Secretion of Matrix Metalloproteinase-9

  • Sung, Ji-Hye;Yoo, Seung-Hee;Park, Hae-Kyung;Chong, Young-Hae
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.1
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    • pp.9-18
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    • 2000
  • Immunological mechanisms involving the release of inflammatory factors by HIV-1 infected microglia in the brain have been implicated in the pathogenesis of HIV dementia (HIVD). Since the regulation of matrix metalloproteinases (MMPs) activity can be influenced by variety of inflammatory mediators, this study was undertaken to look for a correlation between the MMP-9 release and the production of TNF-${\alpha}$ in response to HIV-1 p24 in the human monocyte cell line THP-1 as a model for microglia. First, it was shown that HIV-l core p24 antigen induced THP-1 to secrete MMP-9 in a dose response manner while it elicited a little effect on MMP-2 release in human astroglial cell line T98G. Next, it was found that p24 induced THP-1 to secrete TNF-${\alpha}$ without prior differentiation into macrophages by phorbol myristate acetate (PMA) treatment. Furthermore, anti-TNF-${\alpha}$ neutralizing antibodies significantly blocked p24-induced MMP-9 release in a dose dependent manner. Our data indicate that p24 antigen induces monocytic MMP-9 release by triggering up-regulation of TNF-${\alpha}$ secretion.

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