• Title/Summary/Keyword: rho-O cells

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Repair of UV-induced Cyclobutane Pyrimidine Dimers in Human Mitochonrial DNA-less Cells

  • Ikushima, Takaji;Gu, Ning;Tanizaki, Yuichi
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.479-481
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    • 2002
  • UV-induced DNA damage causes cell killing and mutations leading to carcinogenesis. In normal human cells, UV damage such as cyclobutane pyrimidine dimers (CPDs) and primidine-prymidone (6-4) photoproducts are mainly repaired by nucleotide excision repair mechanism. The molecular processes have been well characterized recently. To know the influence of mitochondrial genome on the nucleotide excision repair mechanism against CPDs, we comparatively examined the production of CPDs by UVC irradiation and their repair kinetics in human cells completely lacking mitochondrial DNA (mtDNA) and the parental HeLa S cells. Whole DNA extracted from the cells exposed to UVC was treated with T4-endonuclease V to break the phosphodiester bond adjacent to CPDs. The DNA was electrophoresed in a denaturing agarose gel, which was visualized by ethidium bromide staining. The relative amount of CPDs was determined by image analysis using NIH Image software. MtDNA- less (rho-O) cells were apparently more sensitive to UVC than HeLa S cells, while the level of induction of CPDs in rho-O and HeLa cells was comparable. The repair of CPDs was less efficient in rho-O cells compared with HeLa cells. The residual amount of CPDs after 24-h repair was larger in rho-O cells than in HeLa cells where more than 90 % of CPDs were repaired by then. The non-repaired CPDs would lead to apoptosis in rho-O cells. These results suggest that mitochondrial genome may contribute to some ATP-dependent steps in nucletide excision repair by supplying sufficient ATP which is generated through a respiratory chain in mitochondria.

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H2O2 Inhibits Proliferation and Mediates Suppression of Migration via DLC1/RhoA Signaling in Cancer Cells

  • Ma, Long;Zhu, Wen-Zhen;Liu, Ting-Ting;Fu, Hui-Ling;Liu, Zhao-Jun;Yang, Bing-Wu;Song, Tai-Yu;Li, Guo-Rong
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.4
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    • pp.1637-1642
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    • 2015
  • Background: RhoGTPase-activating proteins (RhoGAPs) regulate RhoGTPases in cells, but whether individual reactive oxygen species (ROS) regulate RhoGAPs is unknown. Our previous published papers have shown that deleted in liver cancer 1 (DLC1) inhibits cancer cell migration by its RhoGAP activity. The present study was designed to explore the role of $H_2O_2$ in regulation of DLC1. Materials and Methods: We treated cells with $H_2O_2$ for 24h and phenotypic changes were analyzed by MTT, RT-PCR, Western blotting, immunofluorescence staining and wound healing assays. Results: $H_2O_2$ downregulated cyclin D1 and cyclin E to inhibit proliferation, and upregulated BAX to induce apoptosis in MCF-7 cells. Compared with non-tumorigenic cells, $H_2O_2$ increased expression of DLC1 and reduced activity of RhoA in cancer cells. Stress fiber production and migration were also suppressed by $H_2O_2$ in MDA-MB-231 cells. Conclusions: Our study suggests that $H_2O_2$ inhibits proliferation through modulation of cell cycle and apoptosis-related genes, and inhibits migration by decreasing stress fibers via DLC1/RhoA signaling.

Improved Energy Conversion Efficiency of Dye-sensitized Solar Cells Fabricated using Open-ended TiO2 Nanotube Arrays with Scattering Layer

  • Rho, Won-Yeop;Chun, Myeoung-Hwan;Kim, Ho-Sub;Hahn, Yoon-Bong;Suh, Jung Sang;Jun, Bong-Hyun
    • Bulletin of the Korean Chemical Society
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    • v.35 no.4
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    • pp.1165-1168
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    • 2014
  • We prepared dye-sensitized solar cells (DSSCs) with enhanced energy conversion efficiency using open-ended $TiO_2$ nanotube arrays with a $TiO_2$ scattering layer. As compared to closed-ended $TiO_2$ nanotube arrays, the energy conversion efficiency of the open-ended $TiO_2$ nanotube arrays was increased from 5.63% to 5.92%, which is an enhancement of 5.15%. With the $TiO_2$ scattering layer, the energy conversion efficiency was increased from 5.92% to 6.53%, which is an enhancement of 10.30%. After treating the open-ended $TiO_2$ nanotube arrays with $TiCl_4$, the energy conversion efficiency was increased from 6.53% to 6.89%, a 5.51% enhancement, which is attributed to improved light harvesting and increased dye adsorption.

A Comparison of ROCK Inhibitors on Human Bone Marrow-Derived Mesenchymal Stem Cell Differentiation into Neuron-Like Cells

  • Lee, Hyun-Sun;Kim, Kwang-Sei;O, Eun-Ju;Joe, Young-Ae
    • Biomolecules & Therapeutics
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    • v.18 no.4
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    • pp.386-395
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    • 2010
  • Bone marrow-derived mesenchymal stem cells (BM-MSC) are a multipotent cell population that can differentiate into neuron-like cells. Previously it has been reported that murine BM-MSC can differentiate into neuron-like cells by co-treatment with a Rho-associated kinase (ROCK) inhibitor -Y27632 and $CoCl_2$. In this study, we compared several ROCK inhibitors for the ability to induce human BM-MSCs to differentiate into neuron-like cells in the presence of $CoCl_2$. Y27632 with high specificity for ROCK at 1-30 ${\mu}M$ was best at inducing neuronal differentiation of MSCs. Compared to HA1077 and H1152, which also effectively induced morphological change into neuron-like cells, Y27632 showed less toxicity even at 100 ${\mu}M$, and resulted in longer multiple branching processes at a wide range of concentrations at 6 h and 72 h post-induction. H89, however, which has less specificity by inhibition of protein kinase A, S6 kinase 1 and MSK1 with similar or greater potency, was less effective at inducing neuronal differentiation of MSCs. Simvastatin, which can inhibit Rho, Ras, and Rac by blocking the synthesis of isoprenoid intermediates, showed little activity for inducing morphological changes of MSCs into neuron-like cells. Accordingly, the expression patterns for neuronal cell markers,including ${\beta}$-tubulin III, neuron-specific enolase, neurofilament, and microtubule-associated protein, were consistent with the pattern of the morphological changes. The data suggest that the ROCK inhibitors with higher specificity are more effective at inducing neuronal differentiation of MSCs.

Analysis for Regulatory Elements in Yeast MGMT Gene Transcription

  • Joo, Jae-Hoon;Kim, Woo-Jae;Rho, Jae-Kyun;Choe, Jae-Hyun;Choe, Soo-Young;Sang-Dai
    • Animal cells and systems
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    • v.2 no.2
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    • pp.287-295
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    • 1998
  • The Saccharomyces cerevisiae MGMT gene encodes a O6-methylguanine DNA methyltransferase that protects cells from mutation or death by DNA alkylating agents. Using an in vitro transcription system, we analyzed its promoter region to find regulatory elements for transcription initiation. DNase I footprinting and a transcription assay showed that a functional TATA box, 5'-TGATATAGCA-3', is located in the region spanning from -25 to -34. We also found one upstream repressing sequence (URS), -333 to -213, by promoter deletion and competition analysis. Gel mobility shift assays and Southwestern blot analysis using URS region indicate specific complex formations. These results indicate that several cis-acting and trans-acting elements might be involved in the transcriptional regulation of the S. cerevisiae MGMT gene.

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Antidiabetic Activities of Korean Red Pine (Pinus densiflora) Inner Bark Extracts

  • Min, Hee-Jeong;Kim, Eun-Ji;Shinn, Seong-whan;Bae, Young-Soo
    • Journal of the Korean Wood Science and Technology
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    • v.47 no.4
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    • pp.498-508
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    • 2019
  • This study was carried out to investigate the potential of Korean red pine (Pinus densiflora) inner bark extracts as an antidiabetic agent. The ethyl acetate soluble fraction of the bark extracts was chromatographed on a Sephadex LH-20 column to yield five compounds, which structures were elucidated by NMR spectroscopy. The isolated compounds were (+)-catehin, (-)-epicatechin, taxifolin, taxifolin-3'-O-${\beta}$-D-(+)-glucose and $\tilde{n}$-courmaric acid. The antidiabetic activity of the different fractions, including the crude extracts and isolated compounds, was evaluated by ${\beta}$-cells insulin secretion and glucose uptake in skeletal muscle cells. The insulin secretion was 128% for taxifolin at $25{\mu}g/mL$. However, the other samples had no effect on this test. For the glucose uptake activity assay, $1{\mu}M$ insulin and 2 mM metformin were used as controls. Both the crude extract and taxifolin showed relatively low activity values, but the other samples yielded glucose uptake values over 260%. ${\rho}$-courmaric acid showed the highest uptake (270%). The results confirmed that Korean red pine extracts may be used as a hypoglycemic agent.

Reactive Oxygen Species and Cytotoxicity of Bamboo (Phyllostachys pubescens) Sap (대나무수액의 활성산소 소거활성과 세포독성)

  • Cho, Sook-Hyun;Choi, Yong-Jo;Rho, Chi-Woong;Choi, Chul-Yung;Kim, Deok-Song;Cho, Sung-Hwan
    • Food Science and Preservation
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    • v.15 no.1
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    • pp.105-110
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    • 2008
  • The antioxidant properties of bamboos sap isolated from Phyllostachys pubescens were investigated. This product scavenged intracellular reactive oxygen species (ROS) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, and prevented lipid peroxidation. The radical scavenging activity of bamboo sap protected the viability of peritoneal macrophage cells exposed to hydrogen peroxide $(H_2O_2)$, Furthermore, bamboo sap reduced apoptotic cell formation induced by $H_2O_2$ as demonstrated by decreases in the number of hypo-diploid cells am apoptotic cell body formation. These results indicate that bamboo sap has radical scavenging activity and ameliorates $H_2O_2$ induced cytotoxicity.

ISOLATION OF PORCINE MULTIPOTENTIAL SKIN-DERIVED PRECURSOR CELLS AND ITS MULTILINEAGE DIFFERENTIATION (미니돼지에서 다능성 피부유래 전구세포의 추출과 이의 다배엽 세포로의 분화유도에 대한 연구)

  • Choi, Moon-Jeong;Byun, June-Ho;Kang, Eun-Ju;Rho, Gyu-Jin;Kim, Uk-Kyu;Kim, Jong-Ryoul;Park, Bong-Wook
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.34 no.6
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    • pp.588-593
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    • 2008
  • There are increasing reports regarding regeneration of the defected tissues using tissue engineering technique. In this technique, multipotential stem cells are essential. There are many potential sources of adult stem cells, such as bone marrow, umbilical cord blood, fat, muscle, dental tissues and skin. Among them, skin is highly accessible and easily obtained with a minimum of donor site complications. Moreover, skin is an abundant adult stem cell sources and has the potential for self-replication and immune privilege. In this study, we isolated skin-derived precursor cells (SKPs) from the ear of adult miniature pigs. In these SKPs, the expression of transcriptional factors, Oct-4, Sox-2, and Nanog were detected by RT-PCR. In vitro osteogenesis and adipogenesis were observed at 3 weeks after transdifferentiations as assayed by positive von Kossa and Oil-red O staining, respectively. In addition, expression of osteocalcin and osteonectin in the osteogenic differentiation medium and $PPAR{\gamma}2$ and aP2 in the adipogenic differentiation medium were detected by RT-PCR. In vitro neurogenesis of porcine SKPs was observed during 24 and 72 hours after treatment of neurogenic differentiation medium. The results of this study suggest that SKPs demonstrate the properties of pluripotence or multipotence and multi-lineage differentiation. This indicates that autogenous SKPs are a reliable and useful source of adult stem cells for regenerative medicine.

Role of hydrogen peroxide in Rac1 mediated activation of p70s6k signaling pathway

  • Bae, Gyu-Un;Kwon, Hyoung-Keun;Kim, Gwan-Tae;Kim, Yong-Kee;Yoon, Jong-Woo;Cho, Eun-Jung;Lee, Hyang-Woo;Han, Jeung-Whan
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.222.1-222.1
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    • 2003
  • The signal transduction pathway leading to the activation of the p70s6k plays an important role in the progression of cells from G0/G1 to S phase of the cell cycle but remains incompletely characterized. We investigated the role of the Rho family G protein Rac1 in H2O2-mediated p70s6k activation. Transient expression of a dominant negative mutants of the small GTP-binding proteins Rac1 (Rac1N17) and Cdc42(Cdc42N17) showed reduced levels of slower migration on Western blots of one-dimensional SDS-PAGE in p70s6k and ERK1/2 by PDFG stimulation. (omitted)

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