• Title/Summary/Keyword: Potential Interference

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β3GnT8 Regulates Laryngeal Carcinoma Cell Proliferation Via Targeting MMPs/TIMPs and TGF-β1

  • Hua, Dong;Qin, Fang;Shen, Li;Jiang, Zhi;Zou, Shi-Tao;Xu, Lan;Cheng, Zhi-Hong;Wu, Shi-Liang
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.5
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    • pp.2087-2093
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    • 2012
  • Previous evidence showed ${\beta}1$, 3-N-acetylglucosaminyltransferase 8 (${\beta}3GnT8$), which can extend polylactosamine on N-glycans, to be highly expressed in some cancer cell lines and tissues, indicating roles in tumorigenesis. However, so far, the function of ${\beta}3GnT8$ in laryngeal carcinoma has not been characterized. To test any contribution, Hep-2 cells were stably transfected with sense or interference vectors to establish cell lines that overexpressed or were deficient in ${\beta}3GnT8$. Here we showed that cell proliferation was increased in ${\beta}3GnT8$ overexpressed cells but decreased in ${\beta}3GnT8$ knockdown cells using MTT. Furthermore, we demonstrated that change in ${\beta}3GnT8$ expression had significant effects on tumor growth in nude mice.We further provided data suggesting that overexpression of ${\beta}3GnT8$ enhanced the expression of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) at both the mRNA and protein levels, associated with shedding of tissue inhibitors of metalloproteinase TIMP-2. In addition, it caused increased production of transforming growth factor beta 1 (TGF-${\beta}1$), whereas ${\beta}3GnT8$ gene knockdown caused the reverse effect. The results may indicate a novel mechanism by which effects of ${\beta}3GnT8$ in regulating cellular proliferation are mediated, at least in partvia targeting MMPs/TIMPs and TGF-${\beta}1$ in laryngeal carcinoma Hep-2 cells. The finding may lay a foundation for further investigations into the ${\beta}3GnT8$ as a potential target for therapy of laryngeal carcinoma.

Down-regulation of SENP1 Expression Increases Apoptosis of Burkitt Lymphoma Cells

  • Huang, Bin-Bin;Gao, Qing-Mei;Liang, Wei;Xiu, Bing;Zhang, Wen-Jun;Liang, Ai-Bin
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.5
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    • pp.2045-2049
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    • 2012
  • Objective: To investigate the effect of down-regulation of Sentrin/SUMO-specific protease 1 (SENP1) expression on the apoptosis of human Burkitt lymphoma cells (Daudi cells) and potential mechanisms. Methods: Short hairpin RNA (shRNA) targeting SENP1 was designed and synthesized and then cloned into a lentiviral vector. A lentiviral packaging plasmid was used to transfect Daudi cells (sh-SENP1-Daudi group). Daudi cells without transfection (Daudi group) and Daudi cells transfected with blank plasmid (sh-NC-Daudi group) served as control groups. Flow cytometry was performed to screen GFP positive cells and semiquantitative PCR and Western blot assays were employed to detect the inference efficiency. The morphology of cells was observed under a microscope before and after transfection. Fluorescence quantitative PCR and Western blot assays were conducted to measure the mRNA and protein expression of apoptosis related molecules (caspase-3, 8 and 9). After treatment with $COCl_2$ for 24 h, the mRNA and protein expression of hypoxia inducible factor -$1{\alpha}$ (HIF-$1{\alpha}$) was determined. Results: Sequencing showed the expression vectors of shRNA targeting SENP1 to be successfully constructed. Following screening of GFP positive cells by FCM, semiqualitative PCR showed the interference efficiency was $79.2{\pm}0.026%$. At 48 h after transfection, the Daudi cells became shrunken, had irregular edges and presented apoptotic bodies. Western blot assay revealed increase in expression of caspase-3, 8 and 9 with prolongation of transfection (P<0.05). Following hypoxia treatment, mRNA expression of HIF-$1{\alpha}$ remained unchanged in three groups (P>0.05) but the protein expression of HIF-$1{\alpha}$ markedly increased (P<0.05). However, in the sh-SENP1-Daudi group, the protein expression of HIF-$1{\alpha}$ remained unchanged Conclusion: SENP1-shRNA can efficiently inhibit SENP1 expression in Daudi cells. SENP1 inhibition may promote cell apoptosis. These findings suggest that SENP1 may serve as an important target in the gene therapy of Burkitts lymphoma.

Structure and Magnetic Properties of Ho and Ni Co-doped BiFeO3 Ceramics

  • Hwang, J.S.;Yoo, Y.J.;Park, J.S.;Kang, J.H.;Lee, K.H.;Lee, B.W.;Kim, K.W.;Lee, Y.P.
    • Proceedings of the Korean Vacuum Society Conference
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    • 2014.02a
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    • pp.183-183
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    • 2014
  • Recently, multiferroic materials gain much attention due to their fascinating fundamental physical properties. These materials offer wide range of potential applications such as data storage, spintronic devices and sensors, where both electronic and magnetic polarizations can be coupled. Among single-phase multiferroic materials, $BiFeO_3$ is typical because of the room-temperature magnetoelectric coupling in view of long-range magnetic- and ferroelectric-ordering temperatures. However, $BiFeO_3$ is well known to have large leakage current and small spontaneous polarization due to the existence of oxygen vacancies and other defects. Furthermore the magnetic moment of pure $BiFeO_3$ is very weak owing to its antiferromagnetic nature. Recently, various attempts have been performed to improve the multiferroic properties of $BiFeO_3$ through the co-doping at the A and the B sites, by making use of the fact that the intrinsic polarization and magnetization are associated with the lone pair of $Bi^{3+}$ ions at the A sites and the partially-filled 3d orbitals of $Fe^{3+}$ ions at the B sites, respectively. In this study, $BiFeO_3$, $Bi_{0.9}Ho_{0.1}FeO_3$, $BiFe_{0.97}Ni_{0.03}O_3$ and $Bi_{0.9}Ho_{0.1}Fe_{0.97}Ni_{0.03}O_3$ bulk compounds were prepared by solid-state reaction and rapid sintering. High-purity $Bi_2O_3$, $Ho_2O_3$, $Fe_2O_3$ and $NiO_2$ powders with the stoichiometric proportions were mixed, and calcined at $500^{\circ}C$ for 24 h to produce the samples. The samples were immediately put into an oven, which was heated up to $800^{\circ}C$ and sintered in air for 1 h. The crystalline structure of samples was investigated at room temperature by using a Rigaku Miniflex powder diffractometer. The field-dependent and temperature-dependent magnetization measurements were performed with a vibrating-sample magnetometer and superconducting quantum-interference device.

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Quorum Quenching Enzymes and Biofouling Control (정족수 제어효소와 biofouling 제어)

  • Jeon, Young Jae;Jeong, Won-Geom;Heo, Hye-Sook
    • Journal of Life Science
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    • v.26 no.12
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    • pp.1487-1497
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    • 2016
  • Bacterial cell to cell communication strategies called quorum sensing (QS) using small diffusible signaling molecules (auto-inducers) govern the expression of various genes dependent on their population density manner. As a consequence of synthesis and response to the signaling molecules, individual planktonic cells synchronized group behaviors to control a diverse array of phenotypes such as maturation of biofilm, production of extra-polymeric substances (EPS), virulence, bioluminescence and antibiotic production. Many studies indicated that biofilm formations are associated with QS signaling molecules such as acyl-homoserine lactones (AHLs) mainly used by several Gram negative bacteria. The biofilm maturation causes undesirable biomass accumulation in various surface environments anywhere water is present called biofouling, which results in serious eco-technological problems. Numerous molecules that interfere the bacterial QS called quorum quenching (QQ), have been discovered from various microorganisms, and their functions and mechanisms associated with QS have also been elucidated. To resolve biofouling problems related to various industries, the novel approach based on QS interference has been emerged attenuating multi-drug resisting bacteria appearance and environmental toxicities, which may provide potential advantages over the conventional anti-biofouling approaches. Therefore this paper presents recent information related to bacterial quorum sensing system, quorum quenching enzymes that can control the QS signaling, and lastly discuss the anti-biofouling approaches using the quorum quenching.

Measurement of Human Cytochrome P450 Enzyme Induction Based on Mesalazine and Mosapride Citrate Treatments Using a Luminescent Assay

  • Kim, Young-Hoon;Bae, Young-Ji;Kim, Hyung Soo;Cha, Hey-Jin;Yun, Jae-Suk;Shin, Ji-Soon;Seong, Won-Keun;Lee, Yong-Moon;Han, Kyoung-Moon
    • Biomolecules & Therapeutics
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    • v.23 no.5
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    • pp.486-492
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    • 2015
  • Drug metabolism mostly occurs in the liver. Cytochrome P450 (CYP) is a drug-metabolizing enzyme that is responsible for many important drug metabolism reactions. Recently, the US FDA and EU EMA have suggested that CYP enzyme induction can be measured by both enzymatic activity and mRNA expression. However, these experiments are time-consuming and their interassay variability can lead to misinterpretations of the results. To resolve these problems and establish a more powerful method to measure CYP induction, we determined CYP induction by using luminescent assay. Luminescent CYP assays link CYP enzyme activity to firefly luciferase luminescence technology. In this study, we measured the induction of CYP isozymes (1A2, 2B6, 2C9, and 3A4) in cryopreserved human hepatocytes (HMC424, 478, and 493) using a luminometer. We then examined the potential induction abilities (unknown so far) of mesalazine, a drug for colitis, and mosapride citrate, which is used as an antispasmodic drug. The results showed that mesalazine promotes CYP2B6 and 3A4 activities, while mosapride citrate promotes CYP1A2, 2B6, and 3A4 activities. Luminescent CYP assays offer rapid and safe advantages over LC-MS/MS and qRT-PCR methods. Furthermore, luminescent CYP assays decrease the interference between the optical properties of the test compound and the CYP substrates. Therefore, luminescent CYP assays are less labor intensive, rapid, and can be used as robust tools for high-throughput CYP screening during early drug discovery.

Expression of DNA-dependent Protein Kinase and Its Relationship with Epidermal Growth Factor Receptor Signaling in Metastatic Cancer Cell Lines (DNA-PK 및 표피성장인자수용체의 신호전달이 암전이에 미치는 영향)

  • Hwang Jee Young;Kim Sun Hee;Kang Chi Dug;Yoon Man Soo
    • Journal of Life Science
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    • v.15 no.3 s.70
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    • pp.406-414
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    • 2005
  • The genetic instability of cancer cells may be related to inappropriately activated DNA repair pathways. In present study, the modulated expression of DNA-dependent protein kinase (DNA-PK), a major DNA repair protein, in human cancer metastatic cells was tested. The expressions of Ku70/80, regulatory subunit of DNA-PK, and the Ku DNA-binding activity in various highly metastatic cell lines were higher than those in each parental cell line. Also, the expression of DNA-PKcs, catalytic subunit of DNA-PK, and the kinase activity of the whole DNA-PK complex in highly metastatic cells were significantly increased as compared to those of parental cells, suggesting that the enhanced DNA repair capacity of metastatic cells could be associated with aberrant use of DNA repair, which may mediate tumor progression and metastatic potential. Increased EGFR (epidermal growth factor receptor) signaling has been associated with tumor invasion and metastasis, and the linkage between EGFR-mediated signaling and DNA-PK has been suggested. This study showed that PKI166, the new EGFR tyrosine kinase inhibitor, modulated the expressions of Ku70/80 and DNA-PKcs and also revealed the chemosensitization effect of PKI166 against metastatic cells may be in part due to inhibition of Ku70/80. These results suggest that interference in EGFR signaling by EGFR inhibitor resulted in the impairment of DNA repair activity, and thus DNA-PK could be possible molecular targets for therapy against metastatic cancer cells.

Synecological Study of the Forest Vegetation on Mt. Boryeonsan, Chungcheongbuk Province (충청북도 보련산 삼림식생에 대한 군락생태학적 연구)

  • Song, Jong-Suk;Sin, Dong-Guk;Lee, Jang-Soon;Kim, Heon-Kyu;Eom, Goang-Ho
    • Korean Journal of Environment and Ecology
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    • v.23 no.1
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    • pp.66-77
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    • 2009
  • This study aims at classifying and interpreting on the forest vegetation located in the whole area of Mt. Boryeonsan in Noeunmyeon, Chungju city, Chungcheongbuk Province, Korea using the methodology of the ZM school of phytosociology. The forest vegetation was classified into one Larix leptolepis afforestation, five secondary communities including Qurecus acutissima community, Pinus densiflora community, Quercus aliena community, Quercus serrata community, Quercus variabilis community and one association, Ainsliaeo-Quercetum mongolicae Song et al. 1999(Rhododendro schlippenbachii-Quercion mongolicae, Aceri pseudosieboldiani-Quercetalia mongolicae, Quercetea mongolicae). The research expatiated on the relationship between species composition of the forest communities and the environmental conditions, such as human interference, altitude, slope, topography and chemical properties of soil, and also analyzed the life form by each community. As a result, the Ainsliaeo-Quercetum mongolicae was judged as the most stable community among the forest communities identified here. On the other hand, a division of cluster analysis based on the dominance of the component species, appeared similar to the community units classified phytosociologically. Thus, it is inferred that the survey area will be dominated by the potential natural vegetation, Ainsliaeo-Quercetum mongolicae finally, if progressive succession is advanced continuously.

Compact Design and Fabrication of 'Improved QS-MMI' Demultiplexer (Improved QS-MMI' 1.31/1.55μm 파장분리기의 최적화 설계 및 제작)

  • Kim, Nam-Kook;Kim, Jang-Kyum;Choi, Chul-Hyun;O, Beom-Hoan;Lee, Seung-Gol;Park, Se-Gun;Lee, El-Hang
    • Korean Journal of Optics and Photonics
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    • v.16 no.3
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    • pp.248-253
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    • 2005
  • We designed and fabricated a compact multi-mode interference (MMI) wavelength demultiplexer using the concept of 'Improved Quasi-State' modes. The output power and extinction ratio were improved by utilizing modal phase error which is specially occurred in low-index contrast. For a designed demultiplexer, the mode propagation analysis with effective index approximation shows significant improvement of extinction ratio to -25 dB for both $1.31{\mu}m\;and\;1.51{\mu}m$ wavelength region and the split-length was reduced about 1/5 of other MMI devices. The fabricated device shows successful characteristics for both 1.31 and $1.55{\mu}m$ wavelengths. These results demonstrate the potential of low-index materials system and the embossing process for photonic integrated circuits.

Structural basis of novel TRP14, thioredoxin-related protein that regulates TNE-$\alpha$ signaling pathways

  • Woo, Joo-Rang;Jeong, Woo-Jin;Rhee, Sue-Goo;Ryu, Seong-Eon
    • Proceedings of the Korea Crystallographic Association Conference
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    • 2003.05a
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    • pp.18-18
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    • 2003
  • Thioredoxin (Trx) is a small redox protein that is ubiquitously distributed from achaes to human. In diverse organisms, the protein is involved in various physiological roles by acting as electron donor and regulators of transcription and apoptosis as well as antioxidants. Sequences of Trx within various species are 27~69% identical to that of E. coli and all Trx proteins have the same overall fold, which consists of central five β strands surrounded by four α helices. The N-terminal cysteine in WCGPC motif of Trx is redox sensitive and the motif is highly conserved. Compared with general cysteine, the N-terminal cysteine has low pKa value. The result leads to increased reduction activity of protein. Recently, novel thio.edoxin-related protein (TRP14) was found from rat brain. TRP14 acts as disulfide reductase like Trx1, and its redox potential and pKa are similar to those of Trx1. However, TRP14 takes up electrons from cytosolic thioredoxin reductase (TrxR1), not from the mitochondrial thioredoxin reductase (TrxR2). Biological roles of TES14 were reported to be involved in regulating TNF-α induced signaling pathways in different manner with Trx1. In depletion experiments, depletion of TRP14 increased TNF-α induced phosphorylation and degradation of IκBα more than the depletion Trx1 did. It also facilitated activation of JNK and p38 MAP kinase induced by TNF-α. Unlike Trx1, TRP14 shows neither interaction nor interference with ASK1. Here, we determined three-dimensional crystal structure of TRP14 by MAD method at 1.8Å. The structure reveals that the conserved cis-Pro (Pro90) and active site-W-C-X-X-C motif, which may be involved in substrate recognition similar to Trx1 , are located at the beginning position of strand β4 and helix α2, respectively. The TRP14 structure also shows that surface of TRP14 in the vicinity of the active site, which is surrounded by an extended flexible loop and an additional short a helix, is different from that of Trx1. In addition, the structure exhibits that TRP14 interact with a distinct target proteins compared with Trx1 and the binding may depend mainly on hydrophobic and charge interactions. Consequently, the structure supports biological data that the TRP14 is involved in regulating TNF-α induced signaling pathways in different manner with Trx1.

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Characterization of TNNC1 as a Novel Tumor Suppressor of Lung Adenocarcinoma

  • Kim, Suyeon;Kim, Jaewon;Jung, Yeonjoo;Jun, Yukyung;Jung, Yeonhwa;Lee, Hee-Young;Keum, Juhee;Park, Byung Jo;Lee, Jinseon;Kim, Jhingook;Lee, Sanghyuk;Kim, Jaesang
    • Molecules and Cells
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    • v.43 no.7
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    • pp.619-631
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    • 2020
  • In this study, we describe a novel function of TNNC1 (Troponin C1, Slow Skeletal and Cardiac Type), a component of actin-bound troponin, as a tumor suppressor of lung adenocarcinoma (LUAD). First, the expression of TNNC1 was strongly down-regulated in cancer tissues compared to matched normal lung tissues, and down-regulation of TNNC1 was shown to be strongly correlated with increased mortality among LUAD patients. Interestingly, TNNC1 expression was enhanced by suppression of KRAS, and ectopic expression of TNNC1 in turn inhibited KRASG12D-mediated anchorage independent growth of NIH3T3 cells. Consistently, activation of KRAS pathway in LUAD patients was shown to be strongly correlated with down-regulation of TNNC1. In addition, ectopic expression of TNNC1 inhibited colony formation of multiple LUAD cell lines and induced DNA damage, cell cycle arrest and ultimately apoptosis. We further examined potential correlations between expression levels of TNNC1 and various clinical parameters and found that low-level expression is significantly associated with invasiveness of the tumor. Indeed, RNA interference-mediated down-regulation of TNNC1 led to significant enhancement of invasiveness in vitro. Collectively, our data indicate that TNNC1 has a novel function as a tumor suppressor and is targeted for down-regulation by KRAS pathway during the carcinogenesis of LUAD.