• 제목/요약/키워드: Knock control

검색결과 68건 처리시간 0.027초

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

  • 신유섭;고윤우;최은창;강성운;황혜숙;추옥성;이한빈;김철호
    • 대한두경부종양학회지
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    • 제27권1호
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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
    • 통합자연과학논문집
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    • 제12권4호
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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
    • 한국동물생명공학회지
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    • 제36권3호
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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.

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

  • 황성일;정성식;염정국
    • 동력기계공학회지
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    • 제19권2호
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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.

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

  • 임경남;이진범
    • 생명과학회지
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    • 제23권4호
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    • pp.487-494
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    • 2013
  • T-DNA가 표지된 집단에서 AlaAT 유전자가 깨어진 돌연변이체(alaat)를 분리하고, AlaAt1 특이 프라이머를 이용하여 유전자형을 결정하였다. Alaat의 표현형은 대조구와 비교해서 생장의 감소를 보였고, 종자 역시 작고 생산성의 감소를 보였다. 돌연변이체의 AlaAT 활성은 거의 검출되지 않았다. 고염 스트레스 하에서 alaat의 반응을 엽록소 형광과 항산화 효소들의 활성 및 RT-PCR을 이용하여 대조구와 비교하였다. 고염, 건조 및 저온과 같은 모든 비생물적 스트레스에 대한 Fv/Fm은 대조구와 alaat 둘 다 감소를 보였으며, 비생물적 스트레스에 대한 엽록소 형광은 거의 유사하였다. 항산화 효소인 peroxidase (POX)의 활성은 고염 스트레스에 의해 대조구는 증가하나 alaat에서는 오히려 감소하였다. RT-PCR에 의한 cAPX, POX 및 AlaAT mRNA의 수준을 분석한 결과, 효소 활성과 마찬가지로 AlaAt mRNA는 alaat에서 나타나지 않았고, POX2 mRNA는 대조구는 약간의 증가를 보이나 alaat는 거의 검출할 수 없었다. cAPX mRNA는 대조구와 alaat 모두 고염 스트레스에 의해 크게 증가하였다. 이 같은 결과는 AlaAT 유전자 기능의 상실은 염 스트레스 하에서 벼 식물의 생장에 대해 광합성능 보다는 항산화 효소, 특히 POX 활성 및 합성을 변화시킬 수 있음을 제안한다.

A Study on Spark Ignition Natural Gas Engines

  • Cho Haeng-Muk
    • Journal of Advanced Marine Engineering and Technology
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    • 제30권4호
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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
    • 에너지공학
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    • 제19권1호
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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
    • 한국동물생명공학회지
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    • 제34권4호
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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.

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

  • 김도형;배강우;용화심;최은경;김윤섭;박재석;지영구;이계영
    • Tuberculosis and Respiratory Diseases
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    • 제53권3호
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    • pp.275-284
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    • 2002
  • 연구배경: Gemcitabine은 폐암에서 임상적 유용성이 큰 새로운 항암제이다. 저자들은 폐암세포에서 Gemcita bine에 의한 세포 사멸과 p53의 역할을 규명하고자 하였다. 방 법 : 폐암 세포주로 A549와 H358 세포주를 이용하였고 세포 독성 검사는 MTT assay를 이용하였으며 Gemcitabine 농도는 10nM, 100nM, 1uM, 10uM, 100uM을 사용하였다. 세포 주기 검사는 FACScan을 이용하여 분석하였고 p53 활성화 여부는 western blot을 사용하였다. p53 단백질 분해를 촉진시키는 안정적 세포주 A549-E6과 H358-E6을 제조하고 대조 세포주 A549-neo와 H358-neo 세포주와 비교하여 p53의 기능적 knock-out 실험을 시행하였다. p53의 기능적 knock-out은 p53 유도 약제인 doxorubicine 1 M을 사용하여 western blot으로 확인하였다. 결 과 : A549와 H358 세포주에서 Gemcitabine은 농도에 비례한 세포 독성을 보였고 S phase arrest와 p53의 활성화를 유도하였다. 안정적 세포주 A549-E6과 H358-E6은 MTT assay에서 대조 세포주 A549-noo와 H358-noo에 비해 각각 20-30%, 30-40%의 세포 독성 차단효과를 보였다. 결 론 : Gemcitabine은 S phase arrest를 유발시키고 p53 단백질의 활성화를 유도하며 p53의 기능소실이 Gemcitabine에 대한 저항인자로 작용하고 있음을 확인할 수 있었다. 향후 Gemcitabine에 의해 p53의 활성화가 발생하는 신호경로와 p53 활성화에 의한 아포프토시스의 신호경로에 대한 연구가 필요할 것으로 사료된다.

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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    • 제14권11호
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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.