• Title/Summary/Keyword: Knock control

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Analysis of (-)-Epigallocatechin-3-Gallate-Induced Apoptosis and Inhibition of Invasiveness in Oral Cavity Carcinoma Squamous Cell Carcinoma According to Expression of c-Met (구강암편평세포암에서 c-Met 발현여부에 따른 (-)-Epigallocatechin-3-Gallate의 세포사멸 및 종양침습억제효과의 변화분석)

  • Shin, Yoo-Seob;Koh, Yoon-Woo;Choi, Eun-Chang;Kang, Sung-Un;Hwang, Hye-Sook;Choo, Oak-Sung;Lee, Han-Bin;Kim, Chul-Ho
    • Korean Journal of Head & Neck Oncology
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    • v.27 no.1
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    • pp.3-11
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    • 2011
  • Hepatocyte growth factor(HGF) and c-Met play an important role in the control of tumor growth and invasion, and they are known to be good prognostic indicators of patient outcome. Epigallocatechin-3-gallate (EGCG) has been shown to have chemopreventive and therapeutic properties by modulating multiple signal pathways regarding the control of proliferation and invasion of cells. In this study, we evaluated the role of c-Met in EGCG-induced inhibition of invasion and apoptosis in an oral cancer cell line. In KB cells where c-Met was knocked down with siRNA, we performed invasion assay and FACS with Annexin V-FITC/PT staining. In addition, we checked the change of mitochondrial membrane potential(MMP) and the generation of reactive oxygen species(ROS). EGCG-induced inhibition of invasiveness was significantly decreased after the knock-down of c-Met. EGCG-induced apoptosis, MMP change and ROS generation was also reduced in c-Met knock-ed-down KB cells. These results suggest that c-Met is involved in EGCG-induced apoptosis and inhibition of invasiveness of oral cancer cell line.

Studies on the Function of Peroxidasins in Innate Immune System in C. elegans

  • Cho, Jeong Hoon
    • Journal of Integrative Natural Science
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    • v.12 no.4
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    • pp.142-146
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    • 2019
  • Peroxidasin is a unique member of peroxidase family in that it has extracellular matrix (ECM) motif as well as peroxidase activity. Peroxidasins are involved in consolidation the extracellular matrix during development and in innate immune defense. C. elegans has two functional peroxidasins, PXN-1 and PXN-2, and PXN-2 is known to contribute to innate immune system. However, it is not clear of PXN-1 function in innate immune system. Therefore, this study is focused on the function of PXN-1 and the relationship between PXN-1 and PXN-2 in innate defense system in C. elegans. When pxn-1 was knocked down by RNAi, the worm turned to be more resistant to pathogens, Staphylococcus aureus and Pseudomonas aeruginosa and the enhanced resistance was abolished in pxn-1pxn-2 double knock down. By contrast, pxn-2 knock down showed stronger susceptibility to the pathogens. These results suggest that pxn-2 can contribute the pathogen resistance and pxn-1 can suppress the pathogen resistance. To confirm the idea, overexpression experiments were performed. Overexpression of pxn-1 showed more susceptible to pathogens compared to the control and double overexpression of pxn-1pxn-2 overcame the susceptibility of pxn-1 overexpression to the pathogens. On the other hand, pxn-2 overexpression made the worm more resistant to the pathogens and the resistance was maintained in pxn-1pxn-2 double overexpression. The comparison of the susceptibilities to the bacterial pathogens in above mentioned constructs suggests that PXN-1 suppress the function of PXN-2 in defense against bacterial pathogens in Caenorhabditis elegans.

DNA recombinase Rad51 is regulated with UV-induced DNA damage and the DNA mismatch repair inhibitor CdCl2 in HC11 cells

  • You, Hyeong-Ju;Kim, Ga-Yeon;Kim, Seung-Yeon;Kang, Man-Jong
    • Journal of Animal Reproduction and Biotechnology
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    • v.36 no.3
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    • pp.121-128
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    • 2021
  • Increasing the efficiency of HR (homologous recombination) is important for a successful knock-in. Rad51 is mainly involved in homologous recombination and is associated with strand invasion. The HR-related mismatch repair system maintains HR fidelity by heteroduplex rejection and repair. Therefore, the purpose of this study is to control Rad51, which plays a critical role in HR, through UV-induced DNA damage. It is also to confirm the effect on the expression of MMR related genes (Msh2, Msh3, Msh6, Mlh1, Pms2) and HR-related genes closely related to HR through treatment with the MMR inhibitor CdCl2. The mRNA expression of Rad51 gene was confirmed in both HC11 cells and mouse testes, but the mRNA expression of Dmc1 gene was confirmed only in mouse testes. The protein expression of Rad51 and Dmc1 gene increased in UV-irradiated HC11 cells. After 72 hours of treatment with 1 ㎛ of CdCl2, the mRNA expression level of Msh3, Pms2, and Rad51 decreased, but the mRNA expression level of Msh6 and Mlh1 increased in HC11 cells. There was no significant difference in Msh2 mRNA expression between CdCl2 untreated-group and the 72 hours treated group. In conclusion, HR-related gene (Rad51) was increased by UV-induced DNA damage. Treatment of the MMR inhibitor CdCl2 in HC11 cells decreased the mRNA expression of Rad51.

A Study on the Spray and Combustion Characteristics of Direct-injection LPG (직접분사식 LPG의 분무 및 연소 특성에 관한 연구)

  • Hwang, Seong-Ill;Chung, Sung-Sik;Yeom, Jeong-Kuk
    • Journal of Power System Engineering
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    • v.19 no.2
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    • pp.40-48
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    • 2015
  • As advantages of LPG-DI engine, LPG is directly injected into combustion chamber during compression stroke to reduce compression temperature, prevent knock and spontaneous combustion, and adjust engine output using the amount of directly injected fuel, thereby reducing pumping loss caused by throttle valve. Stratified charge can be supplied nearby spark plugs to allow for overall lean combustion, which improves thermal efficiency and can cope with problems regarding emission regulations. In addition, it is characterized by free designing of intake manifold. Despite the fact that LPG-DI has many advantages as described above, there is lack of detailed investigation and study on spray characteristics, combustion flame characteristics, and ignition probability. In this study, a visualization experiment system that consists of visualization combustion chamber, air supply control system, emission control system, LPG fuel supply system, electronic control system and image data acquisition system was designed and manufactured. For supply of stratified charge in the combustion chamber, alignment of injector and spark plugs was made linear.

Salt Stress Responses of an Alanine Aminotransferase Knock-out Mutant of Rice (Oryza sativa L.) (알라닌 아미노기전이효소가 상실된 벼(Oryza sativa L.) 돌연변이체의 고염 스트레스에 대한 반응)

  • Im, Kyoung-Nam;Lee, Chin Bum
    • Journal of Life Science
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    • v.23 no.4
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    • pp.487-494
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    • 2013
  • An AlaAT knock-out mutant (alaat) of rice (Oryza sativa L.) was isolated from T-DNA tagging lines and the genotypes of its progeny were determined with AlaAT1-specific primers. The alaat phenotypes showed decreased growth and grain yield when compared with control plants. The activity of AlaAT1 in the mutant plants was practically undetectable. The responses of alaat plants to growth under salt stress were compared with those of control plants by measuring chlorophyll fluorescence and the activities and mRNA expression of antioxidant enzymes. All abiotic stresses tested (salt, drought, and chilling) caused a similar decrease in chlorophyll fluorescence in both alaat and wild type plants. The activity of peroxidase (POX), an antioxidant enzyme, decreased following salt treatment of alaat plants, while control plant showed an increased activity. The mRNA levels for cAPX (cytosolic ascorbate peroxidase), POX2, and AlaAT were determined by RT-PCR following salt stress. No AlaAT1 mRNA was detected in alaat plants. The POX2 mRNA showed a slightly increased level in the wild type but was not detected in alaat plants, in agreement with the activity assays. The levels of cAPX mRNA were greatly increased in both the wild type and alaat plants. The salt stress effects on rice plant growth are therefore proposed to reflect a loss of function of AlaAT, which alters the activity and synthesis of antioxidant enzymes (especially peroxidases), rather than a direct effect on photosynthesis.

A Study on Spark Ignition Natural Gas Engines

  • Cho Haeng-Muk
    • Journal of Advanced Marine Engineering and Technology
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    • v.30 no.4
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    • pp.455-462
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    • 2006
  • Natural gas is a promising alternative fuel to meet strict engine emission regulations in many countries. Natural gas engines can operate at lean burn and stoichiometric burn conditions with different combustion and emission characteristics. In this paper, the fuel economy, emissions, misfire, knock and cycle-to-cycle variations in indicated mean effective pressure of lean burn natural gas engines are highlighted. Stoichiometric burn natural gas engines are briefly reviewed. To keep the output power and torque of natural gas engines comparable to that of gasoline engines, high boosting pressure should be used. High activity catalyst for methane oxidation and lean deNOx system or three way catalyst with precisely control strategies should be developed to meet stringent emission standards.

Effects of the Intake Valve Timing and the Injection Timing for a Miller Cycle Engine

  • Han, Sung-Bin;Chang, Yong-Hoon;Choi, Gyeung-Ho;Chung, Yon-Jong;Poompipatpong, Chedthawut;Koetniyom, Saiprasit
    • Journal of Energy Engineering
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    • v.19 no.1
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    • pp.32-38
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    • 2010
  • The objective of the research was to study the effects a Miller cycle. The engine was dedicated to natural gas usage by modifying pistons, fuel system and ignition systems. The engine was installed on a dynamometer and attached with various sensors and controllers. Intake valve timing, engine speed, load, injection timing and ignition timing are main parameters. Miller Cycle without supercharging can increase brake thermal efficiency 1.08% and reduce brake specific fuel consumption 4.58%. The injection timing must be synchronous with valve timing, speed and load to control the performances, emissions and knock margin. Throughout these tested speeds, original camshaft is recommended to obtain high volumetric efficiency.

Tight Junction Assembly Ensures Maintenance of Pregnancy during Embryogenesis in a Mouse Model

  • Jeong, Yelin;Choi, Inchul
    • Journal of Animal Reproduction and Biotechnology
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    • v.34 no.4
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    • pp.318-321
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    • 2019
  • Recent studies showed that tight junctions (TJs) integrity and assembly are required for blastocyst development in mouse and pig models. However, the biological functions of TJs associated with embryo implantation and maintenance of pregnancy were not investigated yet. To examine whether disrupted TJs affect further embryo development, we employed RNAi approach and inhibitor treatment. The embryos were injected with Cxadr (Coxsackievirus and adenovirus receptor) siRNA for knock down (KD) and treated with Adam10 (A Disintegrin and Metalloproteinase specific inhibitor 10; GI254023X; SI). We compared blastocyst development and paracellular sealing assay using FITC dextran uptake between control and KD or SI embryos. Finally, we transferred control and Cxadr KD or Adam 10 SI treated blastocyst to uteri of recipients. Cxadr KD and Adam 10 SI showed lower blastocyst development and more permeable to FITC-dextran. Moreover, we observed that half of KD and inhibited embryos failed to maintain pregnancies after the second trimester. Our findings suggested that TJs integrity is required for the maintenance of pregnancy and can be used as a selective marker for the successful application of assisted reproduction technologies.

Gemcitabine-induced Cell Death in Lung Cancer Cells : the Role of p53 (폐암 세포에서 Gemcitabine에 의한 세포 사멸과 p53의 역할)

  • Kim, Doh-Hyung;Bae, Gang-U;Yong, Wha-Shim;Choi, Eun-Kyung;Kim, Youn-Seup;Park, Jae-Seuk;Jee, Young-Koo;Lee, Kye-Young
    • Tuberculosis and Respiratory Diseases
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    • v.53 no.3
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    • pp.275-284
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    • 2002
  • Background : Gemcitabine is a new anti-cancer agent for treating non-small cell lung cancer. Functioning as an antimetabolite, it induces anti-cancer effects by suppressing DNA synthesis after being incorporated into the DNA as a cytosine arabinoside analogue. When Gemcitabine is incorporated into the DNA, the p53 gene may be activated by induction of the DNA defect. However, there are a few studies on the molecular mechanisms of Gemcitabine-induced cell death. This study examined the role of p53 in Gemcitabine-induced cell death. Methods : A549 and NCl-H358 lung cancer cells were used in this study. The cell viability test was done using a MTT assay at Gemcitabine concentrations of 10nM, 100nM, 1uM, 10uM and 100uM. A FACScan analysis with propium iodide staining was used for the cell cycle analysis. Western blot analysis was done to investigate the extent of p53 activation. For the functional knock-out of p53, stable A549-E6 cells and H358-E6 cells were transfected pLXSN-16E6SD which is over expresses the human papilloma virus E6 protein that constantly degrades p53 protein. The functional knock out of p53 was confirmed by Western blot analysis after treatment with a DNA damaging agent, doxorubicine. Results : Gemcitabine exhibited cell toxicity in dose-dependent fashion. The cell cycle analysis resulted in an S phase arrest. Western blot analysis significant p53 activation in time-dependent manner. Gemcitabine-induced cytotoxicity was reduced by 20-30% in the A549-E6 cells and the 30-40% in H358-E6 cells when compared with the A549-neo and H358-neo control cells. Conclusion : Gemcitabine induces an S phase arrest, as expected for the anti-metabolite, and activates the p53 gene, Furthermore, p53 might play an important role in Gemcitabine-induced cell death. Further investigation into the molecular mechanisms on how Gemcitabine activates the p53 gene and its signaling pathway are recommended.

Effects of PTTG Down-regulation on Proliferation and Metastasis of the SCL-1 Cutaneous Squamous Cell Carcinoma Cell Line

  • Xia, Yong-Hua;Li, Min;Fu, Dan-Dan;Xu, Su-Ling;Li, Zhan-Guo;Liu, Dong;Tian, Zhong-Wei
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.11
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    • pp.6245-6248
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    • 2013
  • Aims: To study effects of down-regulation of pituitary tumor-transforming gene (PTTG) on proliferation and metastasis ability of the SCL-1 cutaneous squamous cell carcinoma (CSCC) cell line and explore related mechanisms. Methods: SCL-1 cells were divided into 3 groups (untreated, siRNA control and PTTG siRNA). Cell proliferation assays were performed using a CCK-8 kit and proliferation and metastasis ability were analyzed using Boyden chambers. In addition, expression of MMP-2 and MMP-9 was detected by r-time qPCR and Western blotting. Results: Down-regulation of PTTG could markedly inhibit cell proliferation in SCL-1 cells, compared to untreated and control siRNA groups (P < 0.05). Real-time qPCR demonstrated that expression levels of PTTG, MMP-2 and MMP-9 in the PTTG siRNA group were 0.8%, 23.2% and 21.3% of untreated levels. Western blotting revealed that expression of PTTG, MMP-2 and MMP-9 proteins in the PTTG siRNA group was obviously down-regulated. The numbers of migrating cells ($51.38{\pm}4.71$) in the PTTG siRNA group was obviously lower than that in untreated group ($131.33{\pm}6.12$) and the control siRNA group ($127.72{\pm}5.20$) (P < 0.05), suggesting that decrease of proliferation and metastasis ability mediated by PTTG knock-down may be closely correlated with down-regulation of MMP-2 and MMP-9 expression. Conclusion: Inhibition of PTTG expression may be a new target for therapy of CSCC.