• 제목/요약/키워드: SiHa cell

검색결과 128건 처리시간 0.033초

[Ca2+]-dependent Generation of Intracellular Reactive Oxygen Species Mediates Maitotoxin-induced Cellular Responses in Human Umbilical Vein Endothelial Cells

  • Yi, Sun-Ju;Kim, Kyung Hwan;Choi, Hyun Jung;Yoo, Je Ok;Jung, Hyo-Il;Han, Jeong-A;Kim, Young-Myeong;Suh, In Bum;Ha, Kwon-Soo
    • Molecules and Cells
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    • 제21권1호
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    • pp.121-128
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    • 2006
  • Maitotoxin (MTX) is known as one of the most potent marine toxins involved in Ciguatera poisoning, but intracellular signaling pathways caused by MTX was not fully understood. Thus, we have investigated whether intracellular reactive oxygen species (ROS) are involved in MTX-induced cellular responses in human umbilical vein endothelial cells. MTX induced a dose-dependent increase of intracellular [$Ca^{2+}$]. MTX stimulated the production of intracellular ROS in a dose- and time-dependent manner, which was suppressed by BAPTA-AM, an intracellular $Ca^{2+}$ chelator. Ionomycin also elevated the ROS production in a dose-dependent manner. MTX elevated transamidation activity in a time-dependent manner and the activation was largely inhibited by transfection of tissue transglutaminase siRNA. The activation of tissue transglutaminase and ERK1/2 by MTX was suppressed by BAPTA-AM or ROS scavengers. In addition, MTX-induced cell death was significantly delayed by BAPTA-AM or a ROS scavenger. These results suggest that [$Ca^{2+}$]-dependent generation of intracellular ROS, at least in part, play an important role in MTX-stimulated cellular responses, such as activation of tTGase, ERK phosphorylation, and induction of cell death, in human umbilical vein endothelial cells.

소나무재선충[Bursaphelenchus xylophilus (Steiner and Buhrer) Nickle]의 GaLectin에 대한 특이적인 단클론 항체 제작과 진단에의 활용 (Development of Monoclonal Antibodies Specific to Galectin of Pine Wood Nematode, Bursaphelenchus xylophilus (Steiner and Buhrer) Nickle and Their Utilization for Detection of Pine Wood Nematodes)

  • 김아영;김영하;최보혜;트랑뉴겐;윤경재;이시혁;한혜림;고영호
    • 한국응용곤충학회지
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    • 제57권1호
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    • pp.7-13
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    • 2018
  • 소나무재선충(pine wood nematode, PWN) 감염 소나무를 현장에서 신속하게 진단을 할 수 있는 방법은 현재 없다. 본 연구에서는 PWN특이적인 항원으로 알려진 PWN-GaLectin을 baculovirus발현체계로 발현시켜서 총 1,464개의 fusion hybridoma 세포 주 라이브러리를 제작 하는 항원으로 이용하였다. 총 1,464개의 fusion hybridoma 세포 주 중, PWN-GaLectin에 대한 높은 반응을 보이는 62개의 fusion hybridoma 세포 주를 선별했다. 이들 중, 표준 PWN 감염소나무 PBS추출물과 PWN 단백질 추출물에 강한 반응을 보이는 세포 주 12개를 선별하여 단클론 항체(monoclonal antibody, Mab) 분비세포 주 확립을 위한 limited dilution을 실시하였다. Mab분비세포 주 확립을 위해서 표준 PWN 감염소나무 추출물에 대한 반응이 표준 정상 소나무 추출물 보다 높은 세포 주들을 선별했다. 그리고 추가로 PWN 단백질 추출물에 대해서 3종의 비 병원성 선충 단백질 추출물 보다 높은 반응을 보이는 세포 주들도 선별, 확립했다. 본 연구에서 확립된 Mab들을 우리는 현장과 실험실에서 사용할 수 있는 신속진단키트의 개발에 이용할 수 있을 것이다.

박하에서 분리한 Caffeic Acid Methyl Ester가 자궁경부암 바이러스 발암단백질 E6의 기능에 미치는 영향 (Effects of Caffeic Acid Methyl Ester from Mentha arvensis Linne var. piperascens on Function of E6 Oncoprotein of HPV 16)

  • 백태웅;이경애;안민정;주혜경;오현철;안종석;조용권;명평근;윤도영
    • 약학회지
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    • 제48권6호
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    • pp.328-334
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    • 2004
  • Cervical cancer is one of the leading causes of female death. Viral oncoproteins E6 and E7 are selectively retained and expressed in carcinoma cells infected with HPV (Human pa pilloma virus) type 16 and cooperated in immotalization and transformation of primary keratinocyte. E6 and E7 oncoproteins interfere the functions of tumor suppressor proteins p53 and retinoblasoma protein (pRb), respectively. Among a lots of natural products, Mentha arvensis Linne var.piperascens have inhibitory effects on bindings between E6 oncoprotein and tumor suppressor p53, E3 ubiqutin- protein ligase (E6AP). HPV oncoprotein inhibitors from Mentha piperita L. were isolated by solvent partition and column chromatography (Silica gel, RP-18) and inhibitory compounds were finally purified by HPLC using an ELISA screening system based on binding between E6 and E6AP. The aim of this study is to identify the structure of inhibitory compounds and to investigate whether these compounds have inhibitory effects on functions of E6 oncoprotein. We investigated whether caffeic acid methyl ester (CAM) extracted from Mentha piperita L. could inhibit the function of E6 oncoprotein. CAM inhibited the in vitro binding of E6 and E6AP which are essential for the binding and degradation of the tumor suppressor p53 and also inhibited the proliferation of human cervical cancer cell lines (SiHa and CaSKi) in a dose response manner. These results suggest that CAM inhibited the function of E6 oncoprotein, suggesting that it can be used as a potential drug for the treatment of cervical cancers infected with HPV.

Effect of Gene actA on the Invasion Efficiency of Listeria monocytogenes, as Observed in Healthy and Senescent Intestinal Epithelial Cells

  • Ha, Jimyeong;Oh, Hyemin;Kim, Sejeong;Lee, Jeeyeon;Lee, Soomin;Lee, Heeyoung;Choi, Yukyung;Moon, Sung Sil;Choi, Kyoung-Hee;Yoon, Yohan
    • Journal of Microbiology and Biotechnology
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    • 제28권1호
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    • pp.59-64
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    • 2018
  • Listeria monocytogenes can asymptomatically inhabit the human intestine as a commensal bacterium. However, the mechanism by which L. monocytogenes is able to inhabit the intestine without pathogenic symptoms remains unclear. We compared the invasion efficiency of L. monocytogenes strains with the 268- and 385-bp-long actA gene. Clinical strains SMFM-CI-3 and SMFM-CI-6 with 268-bp actA isolated from patients with listeriosis, and strains SMFM-SI-1 and SMFM-SI-2 with the 385-bp gene isolated from carcasses, were used for inoculum preparation. The invasion efficiency of these strains was evaluated using Caco-2 cells (intestinal epithelial cell line), prepared as normal and healthy cells with tightened tight junctions and senescent cells with loose tight junctions that were loosened by adriamycin treatment. The invasion efficiency of L. monocytogenes strains with the 268-bp-long actA gene was 1.1-2.6-times lower than that of the strains with the 385-bp-long gene in normal and healthy cells. However, the invasion efficiency of both types of strains did not differ in senescent cells. Thus, L. monocytogenes strains with the 268-bp-long actA gene can inhabit the intestine asymptomatically as a commensal bacterium, but they may invade the intestinal epithelial cells and cause listeriosis in senescent cells.

CTRP9 Regulates Growth, Differentiation, and Apoptosis in Human Keratinocytes through TGFβ1-p38-Dependent Pathway

  • Jung, Tae Woo;Park, Hyung Sub;Choi, Geum Hee;Kim, Daehwan;Lee, Taeseung
    • Molecules and Cells
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    • 제40권12호
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    • pp.906-915
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    • 2017
  • Impairment of wound healing is a common problem in individuals with diabetes. Adiponectin, an adipocyte-derived cytokine, has many beneficial effects on metabolic disorders such as diabetes, obesity, hypertension, and dyslipidemia. C1q/TNF-Related Protein 9 (CTRP9), the closest paralog of adiponectin, has been reported to have beneficial effects on wound healing. In the current study, we demonstrate that CTRP9 regulates growth, differentiation, and apoptosis of HaCaT human keratinocytes. We found that CTRP9 augmented expression of transforming growth factor beta 1 ($TGF{\beta}1$) by transcription factor activator protein 1 (AP-1) binding activity and phosphorylation of p38 in a dose-dependent manner. Furthermore, siRNA-mediated suppression of $TGF{\beta}1$ reversed the increase in p38 phosphorylation induced by CTRP9. siRNA-mediated suppression of $TGF{\beta}1$ or p38 significantly abrogated the effects of CTRP9 on cell proliferation and differentiation while inducing apoptosis, implying that CTRP9 stimulates wound recovery through a $TGF{\beta}1$-dependent pathway in keratinocytes. Furthermore, intravenous injection of CTRP9 via tail vein suppressed mRNA expression of Ki67 and involucrin whereas it augmented $TGF{\beta}1$ mRNA expression and caspase 3 activity in skin of type 1 diabetes animal models. In conclusion, our results suggest that CTRP9 has suppressive effects on hyperkeratosis, providing a potentially effective therapeutic strategy for diabetic wounds.

Gravure off-set 인쇄법을 적용한 고효율 다결정 실리콘 태양전지 (Gravure off-set printing method for the high-efficiency multicrystalline-silicon solar cell)

  • 김동주;김정모;배소익;전태현;송하철
    • 한국태양에너지학회:학술대회논문집
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    • 한국태양에너지학회 2011년도 춘계학술발표대회 논문집
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    • pp.293-298
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    • 2011
  • The most widely used method to form an electrode in industrial solar cells are screen printing. Screen printing is characterized by a relatively simple and well-known production sequence with high throughput rates. However the method is difficult to implement a fine line width of high-efficiency solar cells can not be made. The open circuit voltage(Voc) and the short circuit current density(Jsc) and fill factor(FF) need to be further improved to increase the efficiency of silicon solar cells. In this study, gravure offset printing method using the multicrystalline-silicon solar cells were fabricated. Gravure off-set printing method which can print the fine line width of finger electrode can have the ability reduce the shaded area and increase the Jsc. Moreover it can make a high aspect ratio thereby series resistance is reduced and FF is increased. Approximately $50{\mu}m$ line width with $35{\mu}m$ height was achieved. The efficiency of gravure off set was 0.7% higher compare to that of scree printing method.

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골조직 재생 촉진을 위한 Novel Perfusion Bioreactor

  • 이시우;허수진;장지연;정재영;김승언;신정욱
    • 한국재료학회:학술대회논문집
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    • 한국재료학회 2009년도 춘계학술발표대회
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    • pp.46.2-46.2
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    • 2009
  • In vitro 상에서 골조직을 원활하게 재생하기 위해서는 3차원 지지체를 이용한 세포 배양과 세포 배양 시 세포의 형태와 기능을 유지/향상시키기 위한 인체 내 미세 환경 재현은 필수적이다. 따라서 본 연구에서는 뼈 성분과 유사한 생체 활성 물질인 hydroxyapatite (HA)와 생분해성 고분자인 poly $\varepsilon$-caprolactone (PCL)를 복합재료로 이용하여 내부 연결성이 우수한 골조직 재생용 3차원 지지체를 제작하였으며, 골 재생 능력 향상을 위하여 인체내 골조직의 기계적 미세 환경을 체외에서 구현한 새로운 형태의 perfusion bioreactor system을 개발/적용하였다. 또한 본 연구에서 개발된 perfusion bioreactor system의 생물학적 평가를 위해 MG63 (osteoblast like cell, 한국 세포주 은행)과 New Zealand White Rabbit에서 분리한 중간엽 줄기세포를 골조직 재생용 3차원 지지체에 파종하였다. 48시간 동안 안정화 후 perfusion bioreactor system을 이용하여 기계적 자극을 파종된 세포에 인가하였으며, 배양 기간 동안 세포의 증식 확인 및 형태학적 관찰을 실시하였다. 본 연구 결과, perfusion bioreactor system을 이용하여 기계적 자극을 인가한 실험군에서 세포의 증식 및 활성도가 대조군에 비해 우수함을 확인 할 수 있었다. 따라서, perfusion bioreactor를 이용한 세포 배양은 세포의 활성 향상 및 골조직 재생에 도움이 될 것으로 사료된다. 차후 perfusion bioreactor를 이용한 다양한 패턴의 자극이 골재생 능력 및 중간엽 줄기세포의 골 분화능에 미치는 영향에 대한 연구가 필요할 것으로 사료된다.

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HWE에 의한 CdSe 박막의 성장과 광전도 특성 (Growth of CdSe thin films using Hot Wall Eptaxy method and their photoconductive properties)

  • 유상하;홍광준
    • 한국전기전자재료학회:학술대회논문집
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    • 한국전기전자재료학회 2004년도 하계학술대회 논문집 Vol.5 No.1
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    • pp.344-348
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    • 2004
  • The CdSe thin films wee grown on the Si(100) wafers by a hot wall epitaxy method(HWE). The source and substrate temperature are $600^{\circ}C\;and\;430^{\circ}C$ respectively. The crystalline structure of epilayers was investigated by double crystal X-ray diffraction(DCXD). Hall effect on the sample was measured by van der Pauw method and studied on the carrier density and mobility dependence on temperature. From Hall data, the mobility was increased in the timperature range 30K to 150K by impurity scatering and decreased in the temperature range 150K to 293K by the lattice scattering. In order to explore the applicability as a photoconductive cell, we measured the sensitivity($\gamma$), the ratio of photocurrent to darkcurrent(pc/dc), maximum allowable power dissipation(MAPD), spectral response and response time. The results indicated that the photoconductive characteristic were the best for the samples annealed in Cu vapor compare with in Cd, Se, air and vacuum vapour. Then we obtained the sensitivity of 0.99, the value of pc/dc of $1.39{\times}10^7$, the MAPD of 335mV, and the rise and decay time of 10ms and 9.5ms, respectively

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The IRF2BP2-KLF2 axis regulates osteoclast and osteoblast differentiation

  • Kim, Inyoung;Kim, Jung Ha;Kim, Kabsun;Seong, Semun;Kim, Nacksung
    • BMB Reports
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    • 제52권7호
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    • pp.469-474
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    • 2019
  • Kruppel-like factor 2 (KLF2) has been implicated in the regulation of cell proliferation, differentiation, and survival in a variety of cells. Recently, it has been reported that KLF2 regulates the p65-mediated transactivation of $NF-{\kappa}B$. Although the $NF-{\kappa}B$ pathway plays an important role in the differentiation of osteoclasts and osteoblasts, the role of KLF2 in these bone cells has not yet been fully elucidated. In this study, we demonstrated that KLF2 regulates osteoclast and osteoblast differentiation. The overexpression of KLF2 in osteoclast precursor cells inhibited osteoclast differentiation by downregulating c-Fos, NFATc1, and TRAP expression, while KLF2 overexpression in osteoblasts enhanced osteoblast differentiation and function by upregulating Runx2, ALP, and BSP expression. Conversely, the downregulation of KLF2 with KLF2-specific siRNA increased osteoclast differentiation and inhibited osteoblast differentiation. Moreover, the overexpression of interferon regulatory protein 2-binding protein 2 (IRF2BP2), a regulator of KLF2, suppressed osteoclast differentiation and enhanced osteoblast differentiation and function. These effects were reversed by downregulating KLF2. Collectively, our data provide new insights and evidence to suggest that the IRF2BP2/KLF2 axis mediates osteoclast and osteoblast differentiation, thereby affecting bone homeostasis.

Methylation-sensitive high-resolution melting analysis of the USP44 promoter can detect early-stage hepatocellular carcinoma in blood samples

  • Si-Cho, Kim;Jiwon, Kim;Da-Won, Kim;Yanghee, Choi;Kyunghyun, Park;Eun Ju, Cho;Su Jong, Yu;Jeongsil, Kim-Ha;Young-Joon, Kim
    • BMB Reports
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    • 제55권11호
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    • pp.553-558
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    • 2022
  • Hepatocellular carcinoma (HCC) is dangerous cancer that often evades early detection because it is asymptomatic and an effective detection method is lacking. For people with chronic liver inflammation who are at high risk of developing HCC, a sensitive detection method for HCC is needed. In a meta-analysis of The Cancer Genome Atlas pan-cancer methylation database, we identified a CpG island in the USP44 promoter that is methylated specifically in HCC. We developed methylation-sensitive high-resolution melting (MS-HRM) analysis to measure the methylation levels of the USP promoter in cell-free DNA isolated from patients. Our MS-HRM assay correctly identified 40% of patients with early-stage HCC, whereas the α-fetoprotein test, which is currently used to detect HCC, correctly identified only 25% of early-stage HCC patients. These results demonstrate that USP44 MS-HRM analysis is suitable for HCC surveillance.