• 제목/요약/키워드: 2D cell lines

검색결과 353건 처리시간 0.027초

Evaluation of Eye Irritation Potential of Solid Substance with New 3D Reconstructed Human Cornea Model, MCTT HCETM

  • Jang, Won-hee;Jung, Kyoung-mi;Yang, Hye-ri;Lee, Miri;Jung, Haeng-Sun;Lee, Su-Hyon;Park, Miyoung;Lim, Kyung-Min
    • Biomolecules & Therapeutics
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    • 제23권4호
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    • pp.379-385
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    • 2015
  • The eye irritation potential of drug candidates or pharmaceutical ingredients should be evaluated if there is a possibility of ocular exposure. Traditionally, the ocular irritation has been evaluated by the rabbit Draize test. However, rabbit eyes are more sensitive to irritants than human eyes, therefore substantial level of false positives are unavoidable. To resolve this species difference, several three-dimensional human corneal epithelial (HCE) models have been developed as alternative eye irritation test methods. Recently, we introduced a new HCE model, MCTT HCE$^{TM}$ which is reconstructed with non-transformed human corneal cells from limbal tissues. Here, we examined if MCTT HCE$^{TM}$ can be employed to evaluate eye irritation potential of solid substances. Through optimization of washing method and exposure time, treatment time was established as 10 min and washing procedure was set up as 4 times of washing with 10 mL of PBS and shaking in 30 mL of PBS in a beaker. With the established eye irritation test protocol, 11 solid substances (5 non-irritants, 6 irritants) were evaluated which demonstrated an excellent predictive capacity (100% accuracy, 100% specificity and 100% sensitivity). We also compared the performance of our test method with rabbit Draize test results and in vitro cytotoxicity test with 2D human corneal epithelial cell lines.

Anti-Proliferative Activity of Ethanol Extracts from Taxilli Ramulus (Taxillus chinensis (DC.) Danser) Through Cyclin D1 Proteasomal Degradation in Human Colorectal Cancer Cells

  • Park, Gwang Hun;Song, Hun Min;Park, Su Bin;Park, Ji Hye;Shin, Myeong Su;Son, Ho-Jun;Um, Yurry;Jeong, Jin Boo
    • 한국자원식물학회지
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    • 제30권6호
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    • pp.640-646
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    • 2017
  • In this study, we elucidated anti-cancer activity and potential molecular mechanism of 70% ethanol extracts from Taxilli Ramulus (Taxillus chinensis (DC.) Danser) (TR-E70) against human colorectal cancer cells. Anti-cell proliferative effect of TR-E70 was evaluated by MTT assay. The effect of TR-E70 on the expression of cyclin D1 in the protein and mRNA level was evaluated by Western blot and RT-PCR, respectively. TR-E70 suppressed the proliferation of human colorectal cancer cell lines, HCT116 and SW480. Although TR-E70 decreased cyclin D1 expression in protein and mRNA level, decreased level of cyclin D1 protein by TR-E70 more dramatically occurred than that of cyclin D1 mRNA. Cyclin D1 downregulation by TR-E70 was attenuated in presence of MG132. In addition, TR-E70 phosphorylated threonine-286 (T286) of cyclin D1. TR-E70-mediated cyclin D1 degradation was blocked in presence of LiCl as an inhibitor $GSK3{\beta}$ but not PD98059 as an ERK1/2 inhibitor and SB203580 as a p38 inhibitor. Our results suggest that TR-E70 may downregulate cyclin D1 as one of the potential anti-cancer targets through $GSK3{\beta}$-dependent cyclin D1 degradation. From these findings, TR-E70 has potential to be a candidate for the development of chemoprevention or therapeutic agents for human colorectal cancer.

남해 동부 멸치어군의 다중주파수 특성 (The multi-frequency characteristics of anchovy schools in the east of South Sea of South Korea)

  • 박영글;서영일;오택윤;박준성;장충식;강명희
    • 수산해양기술연구
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    • 제51권2호
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    • pp.235-244
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    • 2015
  • The multi-frequency characteristics of anchovy schools were investigated using six acoustic lines collected at 38 and 120 kHz while a primary trawl survey was conducted from 14 April and 18 April of 2014 in off the coast of Tongyeong and Geo-je. Here, the frequency characteristics mean ${\Delta}MVBS$ that is the difference of Mean Volume Backscattering Strength at two frequencies. To use the characteristics effectively, the optimal cell size ($10{\times}2m$) was determined by examining several different cell sizes in consideration with the shapes of fish schools and the ${\Delta}MVBS$ pattern. By examining 6 histograms of ${\Delta}MVBS$, afternoon groups were occupied more in the ${\Delta}MVBS$ range of -6~-4 dB than that of -4~-2 dB, comparing to morning groups. The ${\Delta}MVBS$ range of the morning groups was between -16.9 dB and 11.6 dB, and that of the afternoon groups -16.7 dB and 13.0 dB. The average and standard deviation were $-3.9{\pm}3.6$ dB in the morning and $-4.1{\pm}3.4$ dB in the afternoon, suggesting that morning groups were 2 dB higher than afternoon groups. The ${\Delta}MVBS$ range of all anchovy schools regardless of morning and afternoon was between -16.9 dB and 13.0 dB, their average ${\Delta}MVBS$ was $-4.1{\pm}3.5$ dB. The characteristics can support to identify anchovy species in the waters where multiple fish species are distributed. It is hoped that this study presents the availability and benefit of acoustic data from a primary trawl survey.

Targeting SHCBP1 Inhibits Cell Proliferation in Human Hepatocellular Carcinoma Cells

  • Tao, Han-Chuan;Wang, Hai-Xiao;Dai, Min;Gu, Cheng-Yu;Wang, Qun;Han, Ze-Guang;Cai, Bing
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권10호
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    • pp.5645-5650
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    • 2013
  • Src homology 2 domain containing (SHC) is a proto-oncogene which mediates cell proliferation and carcinogenesis in human carcinomas. Here, the SHC SH2-domain binding protein 1 (SHCBP1) was first established to be up-regulated in human hepatocellular carcinoma (HCC) tissues by array-base comparative genome hybridization (aCGH). Meanwhile, we examine and verify it by quantitative real-time PCR and western blot. Our current data show that SHCBP1 was up-regulated in HCC tissues. Overexpression of SHCBP1 could significantly promote HCC cell proliferation, survival and colony formation in HCC cell lines. Furthermore, knockdown of SHCBP1 induced cell cycle delay and suppressed cell proliferation. Furthermore, SHCBP1 could regulate the expression of activate extracellular signal-regulated kinase 1/2 (ERK1/2) and cyclin D1. Together, our findings indicate that SHCBP1 may contribute to human hepatocellular carcinoma by promoting cell proliferation and may serve as a molecular target of cancer therapy.

온청음(溫淸飮)이 인체 간암세포의 세포주기 G1 Arrest에 미치는 영향 (G1 Arrest of the Cell Cycle by Onchungeum in Human Hepatocarcinoma Cells)

  • 구인모;신흥묵
    • 동의생리병리학회지
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    • 제22권4호
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    • pp.821-828
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    • 2008
  • Onchungeum, a herbal formula, which has been used for treatment of anemia due to bleeding, discharging blood and skin disease. In the present study, it was examined the effects of extract of Onchungeum (OCE) on the growth of human hepatocarcinoma cell lines Hep3B (p53 null type) and HepG2 (p53 wild type) in order to investigate the anti-proliferative mechanism by OCE. Treatment of Hep3B and HepG2 cells to OCE resulted in the growth inhibition in a dose-dependent manner, however Hep3B cell line exhibited a relatively strong anti-proliferative activity to OEC. Flow cytometric analysis revealed that OCE treatment in Hep3B cells caused G1 phase arrest of the cell cycle, which was associated with various morphological changes in a dose-dependent fashion. RT-PCR and immunoblotting data revealed that treatment of OCE caused the down-regulation of cyclin D1 expression, however the levels of cyclin E expression were not changed by OCE. The G1 arrest of the cell cycle was also associated with the induction of Cdk inhibitor p27 by OCE. Because the p53 gene is null in Hep3B cells, it is most likely that the induction of p21 is mediated through a p53-independent pathway. Moreover, p27 detected in anti-Cdk4 and anti-Cdk2 immunoprecipitates from the OCE-treated cells, suggesting that OCE-induced p27 protein blocks Cdk kinase activities by directing binding to the cyclin/Cdk complexes. Furthermore, OCE treatment potently suppresses the phosphorylation of retinoblastoma proteins and the levels of the transcription factor E2F-1 expression. Taken together, these results indicated that the growth inhibitory effect of OCE in Hep3B hepatoma cells was associated with the induction of G1 arrest of the cell cycle through regulation of several major growth regulatory gene products.

Cell Cycle Analysis of Bovine Cultured Somatic Cells by Flow Cytometry

  • H.T. Cheong;D.J. Kwon;Park, J.Y.;J.W. Cho;Y.H. Yang;Park, T.M.;Park, C.K.;B.K. Yang;Kim, C.I.
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2001년도 춘계학술발표대회
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    • pp.69-69
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    • 2001
  • The cell cycle phase in which donor nuclei exist prior to nuclear transfer is an important factor governing developmental rates of reconstituted embryos. It was suggested that quiescent G0 and cycling G1 cells could support normal development of reconstituted embryos. In a quest of optimized donor nuclei treatment prior to nuclear transfer, this study was undertaken to examine the cell cycle characteristics of bovine fetal and adult somatic cells when cultured under a variety of culture treatments and the cell cycle change with the lapse of time after trypsinization. This was archived by measuring the DNA content of cells using flow cytometry, Cultured fetal fibroblast cells, adult skin and muscle cells, and cumulus cells were divided by 3 culture treatments; 1) grown to 60-70% confluency (cycling), 2) serum starved culture, 3) culture to confluency. Trypsinized cells were fixed by 70% ethanol and stained with propidium iodide. For one experiment, trypsinized cells were resuspended in DMEM+10% FBS and incubated for 1.5, 3 and 6 h with occasional shaking before ethanol fixation. Cell cycle phases were determined by flow cytometry enabling calculation of percentages of G0+G1, S and G2+M. The majority of cells were in G0+Gl stage regardless of origin of cells. Cultures that were serum starved or cultured to confluency contained significantly (P<0.05) higher percentages of cells in G0+G1 (89.5-95.4%). For every cell lines and culture treatments, percentages of cells in existing in G0+G1 increased with decreasing of the cell size from large to small. In the serum starved and confluency groups, about 98% of small cells were in G0+G1 Serum starved culture contained higher percentages of small-sized cells (38.5-66.9%) than cycling and confluent cultures regardless of cell lines (P<0.05). After trypsinization of fetal fibroblast and adult skin cells that were serum starved and cultured to confluency, the percentages of cells in G0+G1 significantly increased by incubation for 1.5(95.7-99.5%) and 3.0 h (95.9-98.6%). The results suggest that the efficient synchronization of bovine somatic cells in G0+G1 for nuclear transfer can be established by incubation for a limited time period after trypsinization of serum starved or confluent cells.

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수종의 암세포주에 대한 인삼 사포닌 및 그 분해산물의 구조와 세포독성 관계 (CYTOTOXICITIES OF GINSENG SAPONINS AND THEIR DEGRADATION PRODUCTS AGAINST SOME CANCER CELL LINES AND STRUCTURE-ACTIVITY RELATIONSHIP)

  • 백남인;김신일;이유희;김동선;박종대;이천배
    • 고려인삼학회:학술대회논문집
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    • 고려인삼학회 1993년도 학술대회지
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    • pp.132-137
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    • 1993
  • 한국산 고려 홍삼을 산 또는 알칼리로 가수분해하여, 여러가지 사포게닌과 프로사포게닌을 제조하였으며, 분광학적 데이터와 물리 데이터 등으로부터 이들의 화학 구조를 결정하였다. 이들 중 몇종의 분해산물은 A549, SK - OV. - 3, P388, L1210, SK - Mel - 2 및 K562 등의 암세포에 대하여 세포 독성을 나타내었다. Diol계와 triol계 모두 20번 탄소의 절대구조만이 다른 입체 이성체간의 세포독성의 차이는 인정되지 않았으며, diol 계의 물질들이 triol계 물질보다는 더 높은 활성을 나타내었다. 일반적으로 결합된 탄소의 수가 적을수록 세포독성은 강하여지는 경향??? 보였다.

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In Vivo Antitumor Efficacy of Cw252053, A Folate-based Thymidylate Synthase Inhibitor

  • Oh, Se-Woong;Ha, Jong-Ryul;Baek, Du-Jong
    • Archives of Pharmacal Research
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    • 제24권4호
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    • pp.323-326
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    • 2001
  • Previous studies have demonstrated that CW252053, a quinazoline antifolate, exhibits potent inhibitory activity against thymidylate synthase (TS) as well as cytotoxic activity against tumor cell lines in vitro. In this studys, we evaluated the in vivo antitumor efficacy of CW252053 in the mouse tumor model. Female B6D2F$_1$ mice were injected with LY3.7. 2C TK-/- (thymidine kinase deficient mouse Iymphoma) cells into the gastrocnemius muscle. Then, CW252053 was administered twice daily by intraperitoneal injection for 10 days, and tumor growth was monitored daily by leg diameter measurement. All animals in the vehicle, 5-FU, and low dose (30mgmg/kg CW252053 treated groups died between days 12 and 23 because of the tumor burden. In contrast, dosing with 60 mg/kg of CW252053 produced a cure rat against tumor growth of 37.5% and a survival rate of 50%. Even more significantly, a higher dose of CW252053 (120 mg/kg) elicited both a 100% cure rate and a 100% survival rate at the termination of the study, confirming that this compound has very potent in vivo antitumor activity against tumor growth. During the experimental period of this study no signs of toxicity were observed even at the high CW252053 dosage rate of 120 mg/kg.

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ESD 스프레이를 이용한 OPV 제작 기법 (Fabrication Method of OPV using ESD Spray Coating)

  • 김정수;조정대;김동수
    • 한국신재생에너지학회:학술대회논문집
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    • 한국신재생에너지학회 2010년도 춘계학술대회 초록집
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    • pp.84.2-84.2
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    • 2010
  • PEMS (printed electro-mechanical system) is fabricated by means of various printing technologies. Passive and active components in 2D or 3D such as conducting lines, resistors, capacitors, inductors and TFT, which are printed with functional materials, can be classified in this category. And the issue of PEMS is applied to a R2R process in the manufacturing process. In many electro-devices, the vacuum process is used as the manufacturing process. However, the vacuum process has a problem: it is difficult to apply toa continuous process as a R2R printing process. In this paper, we propose an ESD (electro static deposition) printing process has been used to apply an organic solar cell of thin film forming. ESD is a method of liquid atomization by electrical forces, anelectrostatic atomizer sprays micro-drops from the solution injected into the capillary, with electrostatic force generated by electric potential of about tens of kV. ESD method is usable in the thin film coating process of organic materials and continuous process as a R2R manufacturing process. Therefore, we experiment the thin films forming of PEDOT:PSS layer and Active layer which consist of the P3HT:PCBM. The result of experiment, organic solar cell using ESD thin film coated method is occurred efficiency of about 1.4%. Also, the case of only used to ESD method in the active layer coating is occurred efficiency of about 1.86% as the applying a spin coating in the PEDOT:PSS layer. We can expect that ESD method is possible for continuous process to manufacture in the organic solar cell or OLED device.

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The Role of Stress Granules in the Neuronal Differentiation of Stem Cells

  • Jeong, Sin-Gu;Ohn, Takbum;Jang, Chul Ho;Vijayakumar, Karthikeyan;Cho, Gwang-Won
    • Molecules and Cells
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    • 제43권10호
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    • pp.848-855
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    • 2020
  • Cells assemble stress granules (SGs) to protect their RNAs from exposure to harmful chemical reactions induced by environmental stress. These SGs release RNAs, which resume translation once the stress is relieved. During stem cell differentiation, gene expression is altered to allow cells to adopt various functional and morphological features necessary to differentiate. This process induces stress within a cell, and cells that cannot overcome this stress die. Here, we investigated the role of SGs in the progression of stem cell differentiation. SGs aggregated during the neuronal differentiation of human bone marrow-mesenchymal stem cells, and not in cell lines that could not undergo differentiation. SGs were observed between one and three hours post-induction; RNA translation was restrained at the same time. Immediately after disassembly of SGs, the expression of the neuronal marker neurofilament-M (NF-M) gradually increased. Assembled SGs that persisted in cells were exposed to salubrinal, which inhibited the dephosphorylation of eukaryotic translation initiation factor 2 subunit 1 (eIF2α), and in eIF2α/S51D mutant cells. When eIF2α/S51A mutant cells differentiated, SGs were not assembled. In all experiments, the disruption of SGs was accompanied by delayed NF-M expression and the number of neuronally differentiated cells was decreased. Decreased differentiation was accompanied by decreased cell viability, indicating the necessity of SGs for preventing cell death during neuronal differentiation. Collectively, these results demonstrate the essential role of SGs during the neuronal differentiation of stem cells.