• Title/Summary/Keyword: P19 cell

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Control of X Chromosome Reactivation and Determination of the Ratio of Sex Chromosome to Autosome in Embryonal Carcinoma Cell-Somatic Cell Hybrids (배종양 세포와 체세포 간의 융합 세포에서 X 염색체 재활성화의 조절과 성염색체에 대한 상염색체 비율의 결정)

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    • The Korean Journal of Zoology
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    • v.39 no.1
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    • pp.75-88
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    • 1996
  • OTF9-63 (OTF9) and P19S1O1A1 (P19) embryonal carcinoma (EC) cells were examined for their ability to produce the readivation of inactive X chromosomes from somatic cells. They were hybridized with various somatic cells and resulting HATr EC-somatic cell clones were analysed for their morphology, chromosomal replication pafterns and expression proffies of X-linked and distantiy located genes, Hprt and Pgk-1. The results demonstrated that 0RF9 cells could reactivate the inactive X chromosome whereas P19 cells could not. In adition, EC-somatic cell hybrids tended to reduce the number of sex chromosomes in long-term culture, resulting m 1:2 ratio of sex chromosomes to autosomes The use of EC cell hybrids provides an experimental system for studying the mechanism(s) of the X-reactivatio that is initiated and maintained from meiotic prophase of oogenesis to early embryogenesis.

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The importance of post-thaw subculture for standardizing cellular activity of fresh or cryopreserved mouse embryonic stem cells

  • Ko, Dong Woo;Yoon, Jung Ki;Ahn, Jong il;Lee, Myungook;Yang, Woo Sub;Ahn, Ji Yeon;Lim, Jeong Mook
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.3
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    • pp.335-343
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    • 2018
  • Objective: Remarkable difference in cellular activity was found between early and late subpassaged embryonic stem cell (ESCs) lines, which can be created by subtle changes in cell manipulation protocol. This study subsequently examined whether post-thaw subculture of early subpassaged ESC lines could further affect the activity of the ESCs. Methods: Fresh (as a control treatment) or cryopreserved F1 hybrid (B6CBAF1) early ESC lines (C57BL/6xCBA) of the 4 (P4) or the 19 passage (P19) were subcultured once, twice or six times under the same condition. The post-thaw survival of the ESCs was monitored after the post-treatment subculture and the ability of cell proliferation, reactive oxygen species (ROS) generation, apoptosis and mitochondrial ATP synthesis was subsequently examined. Results: Regardless of the subculture number, P19 ESCs showed better (p<0.05) doubling time and less ATP production than P4 ESCs and such difference was not influenced by fresh or cryopreservation. The difference between P4 and P19 ESC lines became decreased as the post-treatment subculture was increased and the six times subculture eliminated such difference. Similarly, transient but prominent difference in ROS production and apoptotic cell number was detected between P4 and P19 ESCs only at the 1st subculture after treatment, but no statistical differences between two ESC lines was detected in other observations. Conclusion: The results of this study suggest that post-thaw subculture of ESCs under the same environment is recommended for standardizing their cellular activity. The activity of cell proliferation ability and ATP synthesis can be used as parameters for quality control of ESCs.

LncRNA H19/miR-29b-3p/PGRN Axis Promoted Epithelial-Mesenchymal Transition of Colorectal Cancer Cells by Acting on Wnt Signaling

  • Ding, Dayong;Li, Changfeng;Zhao, Tiancheng;Li, Dandan;Yang, Lei;Zhang, Bin
    • Molecules and Cells
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    • v.41 no.5
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    • pp.423-435
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    • 2018
  • This investigation was aimed at working out the combined role of lncRNA H19, miR-29b and Wnt signaling in the development of colorectal cancer (CRC). In the aggregate, 185 CRC tissues and corresponding para-carcinoma tissues were gathered. The human CRC cell lines (i.e. HT29, HCT116, SW480 and SW620) and normal colorectal mucosa cell line (NCM460) were also purchased. Si-H19, si-NC, miR-29b-3p mimics, miR-29b-3p inhibitor, si-PGRN and negative control (NC) were, respectively, transfected into the CRC cells. Luciferase reporter plasmids were prepared to evaluate the transduction activity of $Wnt/{\beta}-catenin$ signaling pathway, and dual-luciferase reporter gene assay was arranged to confirm the targeted relationship between H19 and miR-29b-3p, as well as between miR-29b-3p and PGRN. Finally, the proliferative and invasive capacities of CRC cells were appraised through transwell, MTT and scratch assays. As a result, overexpressed H19 and down-expressed miR-29b-3p displayed close associations with the CRC patients' poor prognosis (P < 0.05). Besides, transfection with si-H19, miR-29b-3p mimic or si-PGRN were correlated with elevated E-cadherin expression, decreased snail and vimentin expressions, as well as less-motivated cell proliferation and cell metastasis (P < 0.05). Moreover, H19 was verified to directly target miR-29b-3p based on the luciferase reporter gene assay (P < 0.05), and miR-29b-3p also bound to PGRN in a direct manner (P < 0.05). Finally, addition of LiCl ($Wnt/{\beta}-catenin$ pathway activator) or XAV93920 ($Wnt/{\beta}-catenin$ pathway inhibitor) would cause remarkably altered E-cadherin, c-Myc, vimentin and snail expressions, as well as significantly changed transcriptional activity of ${\beta}-catenin/Tcf$ reporter plasmid (P < 0.05). In conclusion, the lncRNA H19/miR-29b-3p/PGRN/Wnt axis counted a great deal for seeking appropriate diagnostic biomarkers and treatment targets for CRC.

Influence of the Thickness and Doping Concentration in p- and n-Type Poly-Si Layers on the Efficiency of a Solar Cell Based on a Carbon Fiber

  • Yoon, Min-Seok;Shim, Young Bo;Han, Young-Geun
    • Journal of the Optical Society of Korea
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    • v.19 no.2
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    • pp.199-205
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    • 2015
  • We investigated the effects of the thickness and doping concentration in p- and n-type poly-Si layers on the performance of a solar cell based on a carbon fiber in order to improve the energy conversion efficiency of the cell. The short-circuit current density and open-circuit voltage of the carbon fiber-based solar cell were significantly influenced by the thickness and doping concentration in the p- and n-type poly-Si layers. The solar cell efficiency was successfully enhanced to ~10.5%.

Large Cohort Association of Single Nucleotide Polymorphism of PLA2G4A Gene with White Blood Cell Counts in Korean Population

  • Jung, Suk-Yul
    • Biomedical Science Letters
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    • v.18 no.1
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    • pp.71-75
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    • 2012
  • The PLA2G4A catalyzes the hydrolysis of membrane phospholipids to release arachidonic acid, which is metabolized into lipid-based cellular hormones that regulate inflammatory response. The circulating blood cell numbers can be influenced by stress, infection or inflammation. Quantitative blood cell count traits analysis for the 19 SNPs in the PLA2G4A gene in the Korean Association Resource (KARE) cohort (7551 subjects) was performed. The only one SNP (rs10752979) in the all blood cell count was satisfied with the Bonferroni corrected P-value (<0.00263). Furthermore, 6 of the 19 SNPs in the PLA2G4A gene showed a weak or moderate association with blood cell count (P-values: 0.0048~0.042), suggesting the clue of an association between the PLA2G4A gene and blood cell count, especially white blood cell count. This study may provide insight into the genetic basis of blood cell count related with reaction of infection.

Knockdown of Med19 Suppresses Proliferation and Enhances Chemo-sensitivity to Cisplatin in Non-small Cell Lung Cancer Cells

  • Wei, Ling;Wang, Xing-Wu;Sun, Ju-Jie;Lv, Li-Yan;Xie, Li;Song, Xian-Rang
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.3
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    • pp.875-880
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    • 2015
  • Mediator 19 (Med19) is a component of the mediator complex which is a coactivator for DNA-binding factors that activate transcription via RNA polymerase II. Accumulating evidence has shown that Med19 plays important roles in cancer cell proliferation and tumorigenesis. The involvement of Med19 in sensitivity to the chemotherapeutic agent cisplatin was here investigated. We employed RNA interference to reduce Med19 expression in human non-small cell lung cancer (NSCLC) cell lines and analyzed their phenotypic changes. The results showed that after Med19 siRNA transfection, expression of Med19 mRNA and protein was dramatically reduced (p<0.05). Meanwhile, impaired growth potential, arrested cell cycle at G0/G1 phase and enhanced sensitivity to cisplatin were exhibited. Apoptosis and caspase-3 activity were increased when cells were exposed to Med19 siRNA and/or cisplatin. The present findings suggest that Med19 facilitates tumorigenic properties of NSCLC cells and knockdown of Med19 may be a rational therapeutic tool for lung cancer cisplatin sensitization.

Zinc Restored the Decreased Vascular Smooth Muscle Cell Viability under Atherosclerotic Calcification Conditions

  • Shin, Mee-Young;Kwun, In-Sook
    • Preventive Nutrition and Food Science
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    • v.19 no.4
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    • pp.363-366
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    • 2014
  • Zinc is considered to be involved in maintaining healthy vascular condition. Atherosclerotic calcification of vascular smooth muscle cells (VSMCs) occurs via the mechanism of cell death; therefore, cell viability is a critical factor for preventing VSMC calcification. In this study, we tested whether zinc affected VSMC viability under both normal physiological non-calcifying (0 mM P) and atherosclerotic calcifying conditions (3 and 5 mM P), since VSMC physiological characters change during the VSMC calcification process. The study results showed that an optimal zinc level ($15{\mu}M$) restored the decreased VSMC viability which was induced under low zinc levels (0 and $1{\mu}M$) and calcifying conditions (3 and 5 mM P) at 9 and 15 days culture. This zinc-protecting effect for VSMC viability is more prominent under atherosclerotic calcifying condition (3 and 5 mM P) than normal condition (0 mM P). Also, the increased VSMC viability was consistent with the decreased Ca and P accumulation in VSMC cell layers. The results suggested that zinc could be an effective biomineral for preventing VSMC calcification under atherosclerotic calcifying conditions.

Optimization of Capacitor Threshold VT Implantation for Planar P-MOS DRAM Cell (평면구조 P-MOS DRAM 셀의 커패시터 VT 이온주입의 최적화)

  • Chang Sung-Keun;Kim Youn-Jang
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.19 no.2
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    • pp.126-129
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    • 2006
  • We investigated an optimized condition of the capacitor threshold voltage implantation(capacitor $V_T$ Implant) in planar P-MOS DRAM Cell. Several samples with different condition of the capacitor $V_T$ Implant were prepared. It appeared that for the capacitor $V_T$ Implant of $BF_2\;2.0{\times}l0^{13}\;cm^{-2}$ 15 KeV, refresh time is three times larger than that of the sample, in which capacitor $V_T$ Implant is in $BF_2\;1.0{\times}l0^{13}\;cm^{-2}$ 15 KeV. Raphael simulation revealed that the lowed maximum electric field and lowed minimum depletion capacitance ($C_{MIN}$) under the capacitor resulted in well refresh characteristics.

Protective effect of Oxya chinensis sinuosa methanol extract on UVB-induced damage in human retinal pigment epithelial cells

  • Hyun Jung Lim;Sohyun Park;Joon Ha Lee;In-Woo Kim;HaeYong Kweon;Minchul Seo
    • International Journal of Industrial Entomology and Biomaterials
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    • v.47 no.2
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    • pp.90-98
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    • 2023
  • The human eye, constantly exposed to solar radiation, can be damaged by UV radiation. In particular, ultraviolet B (UVB)-induced damage plays an important role in retinal degeneration and cell aging. In this study, we investigated the protective effects of the methanol extract of Oxya chinensis sinuosa (OCM), an edible insect known for its high protein content (64.2%), and various pharmacological effects, on human retinal pigment epithelial cells. ARPE-19 cells were treated with OCM and subsequently UVB irradiated. Our results showed that OCM effectively attenuates UVB-induced cell damage by reducing MAPK phosphorylation (JNK and p38 MAPK). Additionally, OCM increased the phosphorylation of Akt, and cell cycle regulators, including p21 and p27, in a dose-dependent manner. Moreover, OCM treatment increased ARPE-19 cell proliferation by activating the S6K1/S6 pathway. This study suggests that OCM prevents UVB-induced retinal cell damage by increasing cell proliferation via ROS reduction, suggesting its potential as a functional therapeutic superfood against retinal cell damage.

Methylation Status of H19 Gene in Embryos Produced by Nuclear Transfer of Spermatogonial Stem Cells in Pig

  • Lee, Hyun-Seung;Lee, Sung-Ho;Gupta, Mukesh Kumar;Uhm, Sang-Jun;Lee, Hoon-Taek
    • Reproductive and Developmental Biology
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    • v.35 no.1
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    • pp.67-75
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    • 2011
  • The faulty regulation of imprinting gene lead to the abnormal development of reconstructed embryo after nuclear transfer. However, the correlation between the imprinting status of donor cell and preimplantation stage of embryo development is not yet clear. In this study, to determine this correlation, we used the porcine spermatogonial stem cell (pSSC) and fetal fibroblast (pFF) as donor cells. As the results, the isolated cells with laminin matrix selection strongly expressed the GFR ${\alpha}$-1 and PLZF genes of SSCs specific markers. The pSSCs were maintained to 12 passages and positive for the pluripotent marker including OCT4, SSEA1 and NANOG. The methylation analysis of H19 DMR of pSSCs revealed that the zinc finger protein binding sites CTCF3 of H19 DMRs displayed an androgenic imprinting pattern (92.7%). Also, to investigate the reprogramming potential of pSSCs as donor cell, we compared the development rate and methylation status of H19 gene between the reconstructed embryos from pFF and pSSC. This result showed no significant differences of the development rate between the pFFs ($11.2{\pm}0.8%$) and SSCs ($13.3{\pm}1.1%$). However, interestingly, while the CTCF3 methylation status of pFF-NT blastocyst was decreased (36.3%), and the CTCF3 methylation status of pSSC-NT blastocyst was maintained. Therefore, this result suggested that the genomic imprinting status of pSSCs is more effective than that of normal somatic cells for the normal development because the maintenance of imprinting pattern is very important in early embryo stage.