• Title/Summary/Keyword: FAS1

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Evaluation of Cytotoxicity for Immunity Rejection of US11, hDAF and FasL Transgene-Transfected Cells

  • Kang, Jung Won;Shin, Hyeon Yeong;Oqani, Reza K.;Lin, Tao;Lee, Jae Eun;Kim, So Yeon;Lee, Joo Bin;Jin, Dong Il
    • Reproductive and Developmental Biology
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    • v.41 no.3
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    • pp.57-63
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    • 2017
  • Xenotransplantation is proposed as a solution to the problem of organ shortage. However, transplantation of xenogeneic organs induces an antigen-antibody reaction in ${\alpha}$-1,3-gal structure that are not present in humans and primates, and thus complement is also activated and organs die within minutes or hours. In this study, we used FasL gene, which is involved in the immune response of NK cell, and US11, which suppresses MHC Class I cell membrane surface expression, to inhibit cell mediated rejection in the interspecific immunity rejection, and also hDAF(CD55) was introduced to confirm the response to C3 complement. These genes were tranfeced into Korean native pig fetal fibroblasts using pCAGGS vector. And cytotoxicity of NK cell and human complement was confirmed in each cell line. The US11 inhibited the cytotoxicity of NK cell and, in addition, the simultaneous expression of US11 and Fas ligand showed excellent suppress to T-lymphocyte cytotoxicity, hDAF showed weak resistance to cytotoxicity of natural killer cell but not in CD8+ CTLs. Cytotoxicity study with human complement showed that hDAF was effective for reducing complement reaction. In this studies have demonstrated that each gene is effective in reducing immune rejection.

Efficient Killing Effect of Osteosarcoma Cells by Cinobufacini and Cisplatin in Combination

  • Huang, Tao;Gong, Wei-Hua;Li, Xiu-Cheng;Zou, Chun-Ping;Jiang, Guang-Jian;Li, Xu-Hui;Qian, Hao
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.6
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    • pp.2847-2851
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    • 2012
  • Purpose: To study the killing effects on osteosarcoma cells of cinobufacini and cisplatin in combination and the related mechanisms so as to explore the chemotherapeutic method with integrated traditional Chinese and Western medicines. Methods: Cinobufacini and cisplatin were applied to OS732 cells singly or jointly and survival rates were measured by MTT assay. Changes in cellular shape were observed with inverted phase contrast and fluorescence microscopy and apoptosis rates were analyzed with flow cytometry (FCM). Immunocytochemistry were used to examine the Fas expression of OS732 cells. Results: The combination of cinobufacini and cisplatin had the effect of up-regulating Fas expression and inducing apoptosis. The survival rate of combined application of 100 ${\mu}g/ml$ cinobufacini and 1 ${\mu}g/ml$ cisplatin on OS-732 cells was significantly lower than with either of the agents alone (p<0.01). Changes in cellular shape and apoptotic rates also indicated the apoptosis-inducing effects of combined application were much enhanced. Conclusion: The combination of cinobufacini and cisplatin demonstrated strong killing effects on OS-732 cells which might be related to up-regulation of Fas expression.

H9 Induces Apoptosis via the Intrinsic Pathway in Non-Small-Cell Lung Cancer A549 Cells

  • Kwon, Sae-Bom;Kim, Min-Je;Sun Young, Ham;Park, Ga Wan;Choi, Kang-Duk;Jung, Seung Hyun;Do-Young, Yoon
    • Journal of Microbiology and Biotechnology
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    • v.25 no.3
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    • pp.343-352
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    • 2015
  • H9 is an ethanol extract prepared from nine traditional/medicinal herbs. This study was focused on the anticancer effect of H9 in non-small-cell lung cancer cells. The effects of H9 on cell viability, apoptosis, mitochondrial membrane potential (MMP; ${\Delta}\psi_{m}$), and apoptosisrelated protein expression were investigated in A549 human lung cancer cells. In this study, H9-induced apoptosis was confirmed by propidium iodide staining, expression levels of mRNA were determined by reverse transcriptase polymerase chain reaction, protein expression levels were checked by western blot analysis, and MMP (${\Delta}\psi_{m}$) was measured by JC-1 staining. Our results indicated that H9 decreased the viability of A549 cells and induced cell morphological changes in a dose-dependent manner. H9 also altered expression levels of molecules involved in the intrinsic signaling pathway. H9 inhibited Bcl-xL expression, whereas Bax expression was enhanced and cytochrome C was released. Furthermore, H9 treatment led to the activation of caspase-3/caspase-9 and proteolytic cleavage of poly(ADP-ribose) polymerase; the MMP was collapsed by H9. However, the expression levels of extrinsic pathway molecules such as Fas/FasL, TRAIL/TRAIL-R, DR5, and Fas-associated death receptor were downregulated by H9. These results indicated that H9 inhibited proliferation and induced apoptosis by activating intrinsic pathways but not extrinsic pathways in human lung cancer cells. Our results suggest that H9 can be used as an alternative remedy for human non-small-cell lung cancer.

Induction of Apoptosis by Cisplatin, Heptaplatin and Sunpla in Human Melanoma (SK-MEL-28) Cell Line (인체 흑색종 세포(SK-MEL-28 Cell Line)에서 Cisplatin, Heptaplatin, 그리고 Sulpla에 의한 Apoptosis의 유도)

  • 최수라;명평근
    • YAKHAK HOEJI
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    • v.48 no.2
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    • pp.147-152
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    • 2004
  • A wide variety of cancer chemotherapeutic agents have been shown to induce programmed cell death (PCD, APOPTOSIS) in various tumor cell lines in vitro. cis-Malonato [(4R,5R)-4,5-bis(aminomethyl)-2-isoprpopyl-1,3-dioxolane] platinum(II) (heptaplatin), which is a new drug approved by KFDA in 1999, in a novel platinum-based antitumor agent with clinical potential against stomach cancer and the 3rd generation of the cisplatin. This study was performed to know how heptaplatin and cisplatin and sunpla (mixture of heptaplatin and mannitol) affect on SK-MEL-28 cell line, and how they induce the apoptosis. At EM analysis, the morphology of the cell was changed by treatment of the cisplatin, heptaplatin and sunpla. Apoptotic body formed around plasma membrane, and chromatin condensation represented in nucleus. This phenomenon is one of the characteristic of the apoptosis. The DNA of SK-MEL-28 cell line truncated by cisplatin and sunpla treatment was identified on 2% agarose gel electrophoresis. TUNEL assay was performed to know whether SK-MEL-28 cell die as apoptosis or necrosis by cisplatin, heptaplatin and sunpla. At this result, fluorescence intensity increased according to increase of time and concentration. Therefore, it was identified that cislatin, heptaplatin and sunpla induced apoptosis. Fas expressed on SK-MEL-28 cell membrane by cisplatin, heptaplatin and sunpla was identified by using flow cytometer and the expression of bcl-2(anti-apoptotic gene) decreased according to increase of concentration of the cisplatin, heptaplatin and sunpla. Cisplatin, heptaplatin and sunpla induced apoptosis against SK-MEL-28 cell line, and the apoptotic mechanism was identified as Fas-mediated apoptosis and decreased bcl-2 expression.

Effects of Piperine on Insulin Resistance and Lipid Accumulation in Palmitate-treated HepG2 Cells (Palmitate처리된 인간 간세포주 HepG2 세포에서 piperine의 지질 축적과 인슐린 저항성 기전에 대한 연구)

  • Jung, Hee Jin;Bang, EunJin;Jeong, Seong Ho;Kim, Byeong Moo;Chung, Hae Young
    • Journal of Life Science
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    • v.29 no.9
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    • pp.964-971
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    • 2019
  • Hepatic lipid accumulation and insulin resistance increases in patients with non-alcoholic fatty liver disease. Piperine is a major compound found in black pepper (Piper nigrum) and long pepper (P. longum). Piperine has been used in fine chemical for its anti-cancer, anti-obesity, anti-diabetic, anti-inflammatory and anti-oxidant properties. However, the signaling-based mechanism of piperine and its role as an inhibitor of lipogenesis and insulin resistance in human hepatocyte cells remains ill-defined. In the present study, we explored the effects of piperine on lipid accumulation and insulin resistance, and explored the potential underlying molecular mechanisms in palmitate-treated HepG2 cells. Piperine treatment resulted in a significant reduction of triglyceride content. Furthermore, piperine treatment decreased palmitate-treated intracellular lipid deposition by inhibiting the lipogenic target genes, sterol-regulatory-element-binding protein 1c (SREBP-1c) and fatty acid synthase (FAS); whereas the expression of carnitine palmitoyl transferase (CPT-1) and phosphorylation of acetyl coenzyme A carboxylase (ACC) gene involved in fatty acid oxidation was increased. Moreover, piperine also inhibited the phosphorylation of insulin receptor substrate (IRS)-1 (Ser307). Piperine treatment modulated palmitate-treated lipid accumulation and insulin resistance in HepG2 cells with concomitant reduction of lipogenic target genes, such as SREBP-1 and FAS, and induction of CPT-1-ACC and phosphorylation of IRS-1 (Tyr632)-Akt pathways. Therefore, piperine represents a promising treatment for the prevention of lipid accumulation and insulin resistance.

Dynamic Sliding Behavior of Water Droplets on the Coated Hydrophobic Surfaces (발수코팅된 표면에서의 수적의 동적 전락거동)

  • Song, Jeong-Hwan;Nakajima, Akira
    • Korean Journal of Materials Research
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    • v.17 no.11
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    • pp.569-573
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    • 2007
  • The static and dynamic hydrophobicities of the water droplets placed on a hydrophobic surface coated using a fluoroalkylsilanes monolayer with different molecular chain lengths were investigated through direct observation of the actual droplet motion during the sliding process. The surface roughness of both was found to be less than 1 nm. The static contact angles of the coated FAS-3 and FAS-17 were respectively $80^{\circ}$ and $108^{\circ}$ at $150^{\circ}C$, 1 h. The slope of sliding acceleration against the water droplet mass exhibited an inflection point, thus suggesting the switching of the dominant sliding mode from slipping to rolling. While their sliding angles were similar in value, notable differences were exhibited in terms of their sliding behavior. This can be understood as being due to the contribution of the shear stress difference at the interface between the solid surface and water during the sliding process. These results show that the sliding acceleration of the water droplets depends strongly on the balance between gravitational and retentive forces on the hydrophobic surface.

Full-length Fas-associated Death Domain Protein Interacts with Short Form of Cellular FLICE Inhibitory Protein

  • Jeong, Mi-Suk;Jang, Se-Bok
    • Bulletin of the Korean Chemical Society
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    • v.27 no.1
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    • pp.87-92
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    • 2006
  • Fas-associated death domain protein (FADD) recruits and activates procaspase-8 through interactions between the death effector domains of these two proteins. Cellular FLICE-inhibitory protein (c-FLIP) was identified as a molecule with sequence homology to caspase-8. It has been postulated that c-FLIP prevents formation of the competent death-inducing signaling complex in a ligand-dependent manner, through its interaction with FADD and/or caspase-8. However, the interaction of FADD and $c-FLIP_s$ (short form) in apoptosis signaling has been controversially discussed. We show the purification and the characterization of human full-length FADD and $c-FLIP_s$ expressed in Escherichia coli. The purified FADD and $c-FLIP_s$ are shown as homogeneity, respectively, in SDS-PAGE analysis and light-scattering measurements. The folding properties of the $\alpha$-helical structure of FADD and the super-secondary structure of $c-FLIP_s$ proteins were characterized by circular dichroism spectroscopy. Furthermore, we report here a series of biochemical and biophysical data for FADD-$c-FLIP_s$ binding in vitro. The binding of both FADD and $c-FLIP_s$ proteins was detected by BIAcore biosensor, fluorescence measurement, and size-exclusion column (SEC).

Changes in Membrane Fatty Acid Composition during Entry of Vibrio vulnificus into the Viable But Nonculturable State

  • Day, Ashley P.;Oliver, James D.
    • Journal of Microbiology
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    • v.42 no.2
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    • pp.69-73
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    • 2004
  • Vibrio vulnificus, a Gram-negative bacterium found in estuarine waters, is responsible for over 95% of all seafood-related deaths in the United States. As a result of a temperature downshift to 5$^{\circ}C$, this organism enters the viable but nonculturable (VBNC) state. Changes in the membrane fatty acid (FA) composition of V. vulnificus may be a contributing factor to the ability of this organism to enter into and survive in the VBNC state. This hypothesis was tested by incubating the organism at 5$^{\circ}C$ in arti-ficial sea water and analyzing the cells' FAs during the initial hours of temperature and nutrient down-shift. Prior to downshift, the predominant FAs were 16:0, 16:1 and 18:0. During the first four hours of downshift, statistically significant changes occurred in 15:0, 16:1, 16:0, 17:0, and 18:0. These results indicate that changes in FA composition occur prior to entry of V. vulnificus into the VBNC state, suggesting that the ability to maintain membrane fluidity may be a factor in this physiological response. Cells in which fatty acid synthesis was inhibited did not survive, indicating that active fatty acid metab-olism is essential for entry of cells into the VBNC state.

Peptide Inhibitor for Human Immunodeficiency Virus Type 1 (HIV-1) Protease from a Thermolysin Hydrolysate of Oyster Proteins

  • Lee, Tae-Gee
    • Fisheries and Aquatic Sciences
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    • v.13 no.1
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    • pp.84-87
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    • 2010
  • A peptide that inhibits HIV-1 protease was isolated from a hydrolysate of oyster (Crassostrea gigas) proteins digested with thermolysin. The peptide was using membrane filtration, gel permeation chromatography, ion exchange chromatography, and reverse-phase high performance liquid chromatography. Amino acid sequence of the peptide was determined to be Val-Phe-Glu-Leu. Chemically synthesized Val-Phe-Glu-Leu showed an $IC_{50}$ value of 106 ${\mu}M$.

Effects of UV Sterilization on Quality of Acer mono Sap (자외선 살균이 고로쇠 수액의 품질에 미치는 영향)

  • Choe, Su Bin;Han, Sang Jin;Han, Ouk Hee;Kim, Hak Su;Kang, Sung Tae
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.1
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    • pp.148-152
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    • 2013
  • An ultraviolet (UV) sterilization system was developed to decrease the number of microorganisms in filtered Acer mono sap (FAS). The Acer mono sap (AS) was passed through 18 strips of PTA fluoroplastic tubing with 30 UV lamps (total 1,170 W). During passage, the AS was effectively exposed to the UV rays. The total bacteria, coliform group and fungi were sterilized at a flow rate ranging from 852 to 1,358 mL/min and a UV power higher than 156 W. Although the sensory score of the UV-sterilized AS was significantly lower than that of the untreated AS, the sensory score was better under the condition of 390 W and 852~1,358 mL/min than under the other conditions examined. ANOVA did not reveal a significant difference in pH, total acidity, sugar content and color characteristic under all conditions tested (p<0.05). Considering the overall attributes of the AS quality, the optimum sterilization condition was determined to be 390 W and 852~1,358 mL/min.