• Title/Summary/Keyword: biological cells

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Pathway Analysis in HEK 293T Cells Overexpressing HIV-1 Tat and Nucleocapsid

  • Lee, Min-Joo;Park, Jong-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.19 no.10
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    • pp.1103-1108
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    • 2009
  • The human immunodeficiency virus (HIV)-l protein Tat acts as a transcription transactivator that stimulates expression of the infected viral genome. It is released from infected cells and can similarly affect neighboring cells. The nucleocapsid is an important protein that has a related significant role in early mRNA expression, and which contributes to the rapid viral replication that occurs during HIV-1 infection. To investigate the interaction between the Tat and nucleocapsid proteins, we utilized cDNA micro arrays using pTat and flag NC cotransfection in HEK 293T cells and reverse transcription-polymerase chain reaction to validate the micro array data. Four upregulated genes and nine downregulated genes were selected as candidate genes. Gene ontology analysis was conducted to define the biological process of the input genes. A proteomic approach using PathwayStudio determined the relationship between Tat and nucleocapsid; two automatically built pathways represented the interactions between the upregulated and downregulated genes. The results indicate that the up- and downregulated genes regulate HIV-1 replication and proliferation, and viral entry.

Estudy the Effect of Breast Cancer on Tlr2 Expression in Nb4 Cell

  • Amirfakhri, Siamak;Salimi, Arsalan;Fernandez, Nelson
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.18
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    • pp.8445-8450
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    • 2016
  • Background: Breast cancer is the most common neoplasm in women and the most frequent cause of death in those between 35 and 55 years of age. All multicellular organisms have an innate immune system, whereas the adaptive or 'acquired' immune system is restricted to vertebrates. This study focused on the effect of conditioned medium isolated from cultured breast cancer cells on NB4 neutrophil-like cells. Materials and Methods: In the current study neutrophil-like NB4 cells were incubated with MCF-7 cell-conditioned medium. After 6 h incubation the intracellular receptor TLR2, was analyzed. Results: The results revealed that MCF-7 cell-conditioned medium elicited expression of TLR2 in NB4 cells. Conclusions: This treatment would result in the production of particular stimulants (i.e. soluble cytokines), eliciting the expression of immune system receptors. Furthermore, the flow cytometry results demonstrated that MCF-7 cell-conditioned medium elicited an effect on TLR2 intracellular receptors.

Enzyme-Linked, Biotin-Streptavidin Bacterial-Adhesion Assay for Helicobacter pylori Lectin-Like Interactions with Cultured Cells

  • Murillo, Guzman;Antonia, Maria;Ascencio, Felipe
    • Journal of Microbiology and Biotechnology
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    • v.11 no.1
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    • pp.35-39
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    • 2001
  • A simple method for studying the lectin-like interactions between Helicobacter pylori and cultured human epithelial cell lines was developed using an enzyme-linked, biotin-streptavidin bacterial-adhesion assay. The present study suggests that this method is suitable for evaluating the participation of lectin interactions in the adhesion of H. pylori to cultured HeLa S3 and Kato III cells, both fixed and glycosidase-treated cells, as well as assessing glycoconjugated binding inhibition studies. The time-course and dose-dependent kinetics of the biotin-labeled H. pylori adhesion th the formaldehyde-fixed Hela S3 and Kato III cell lines exhibited saturation. In addition, the binding of the biotin-labeled H. pylori to the formaldehyde-fixed cultured cells was partially blocked by pre-incubation with glycoconjugates and polyclonal antibodies against a heparan sulfate binding protein from H. pylori.

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Protein Interaction Mapping of Translational Regulators Affecting Expression of the Critical Stem Cell Factor Nos

  • Malik, Sumira;Jang, Wijeong;Kim, Changsoo
    • Development and Reproduction
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    • v.21 no.4
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    • pp.449-456
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    • 2017
  • The germline stem cells of the Drosophila ovary continuously produce eggs throughout the life-span. Intricate regulation of stemness and differentiation is critical to this continuous production. The translational regulator Nos is an intrinsic factor that is required for maintenance of stemness in germline stem cells. Nos expression is reduced in differentiating cells at the post-transcriptional level by diverse translational regulators. However, molecular mechanisms underlying Nos repression are not completely understood. Through three distinct protein-protein interaction experiments, we identified specific molecular interactions between translational regulators involved in Nos repression. Our findings suggest a model in which protein complexes assemble on the 3' untranslated region of Nos mRNA in order to regulate Nos expression at the post-transcriptional level.

Protein Profile of Embryonic Stem Cells Following Differentiation

  • D. I. Jin;Kang, H. S;Kim, H. J.;Lee, S. H.;Park, C. S.;K. S. Im;Lee, H. M.
    • Korean Journal of Animal Reproduction
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    • v.26 no.4
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    • pp.377-384
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    • 2002
  • To investigate the expression patterns of proteins and growth factor signals in differentiated rabbit embryonic stem (ES) cells, ES cells with confluent stage grown of feeder layer and differentiated cells into embryoid bodies (EB) without feeder cell were applied to protein gel and Western blotting analysis. There were 66kDa and 28kDa specifically expressed in differentiated ES cell but not in undifferentiated ES cell while 25kDa protein band showed up in only undifferentiated ES cells. Also there were some difference of protein bands in several area of gel between differentiated and undifferentiated ES cells such as about 100 kDa, 50kDa and 27kDa areas, but there was no difference in band pattern of one-dimensional gel analysis between mouse ES cells and rabbit ES cells. IGF-I receptor and EGF receptor were expressed in differentiated cells and undifferentiated cells. And ICF-I and EGF were not expressed in both differentiated and undifferentiated cells. These results indicated that ES cells express their own proteins to inhibit differentiation while EB cells synthesize different proteins to differentiate, and 16F-I receptor and EGF receptor were expressed in both ES and EB cells probably for the different functions.

Microfluidic Devices for Cell Analysis

  • Bachman, Mark;Li, G.P.
    • Proceedings of the Materials Research Society of Korea Conference
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    • 2009.11a
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    • pp.3.2-3.2
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    • 2009
  • Microfluidics and BioMEMStechnology has increasingly been used as a tool for studying small volumes oftissue and even individual cells. One of the most important benefits ofmicrofluidic technology is the potential to build devices that analyze and sortmammalian cells. The "sorting problem" typically requires that a fewcells be selected and isolated from a larger population of hundreds, thousandsor even millions of other cells. For example, cancer tumor cells may resideamong a large population of healthy cells, but it would be of great interest toidentify, isolate and study only the cancer cells. In another application, onemay want to determine the number of white blood cells within a sample of blood.We have developed microfluidic devices that enable researchers to select cellsfrom a population by a variety of methods, including antibody staining,dielectrophoretic selection, and physical size selection. These devices haveapplications in cancer research where cancer cells must be identified fromnormal tissue, but where only small samples of tissue are available. In thistalk, we will present some of our microfluidic cell sorting devices, discusstheir physical principles, and their use in biological applications.

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Induction of apoptosis by water extract Glycyrrhizae radix in human bladder T24 cancer cells (인체 방광암 T24 세포에서 Glycyrrhizae radix 열수추출물에 의한 apoptosis 유도)

  • Eom, Jung Hye;Hwang, Buyng Su;Jeong, Yong Tae;Kim, Min-Jin;Shin, Su Young;Kim, Chul Hwan;Lee, Seung Young;Choi, Kyung Min;Cho, Pyo Yun;Jeong, Jin-Woo;Oh, Young Taek
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.04a
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    • pp.111-111
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    • 2019
  • Glycyrrhizae radix is one of the most frequently prescribed ingredients in Oriental medicine, and G. radix extract has been shown to exert anti-cancer effects. However, the cellular and molecular mechanisms of apoptosis by G. radix are poorly defined. In the present study, it was examined the biochemical mechanisms of apoptosis by water extract of G. radix (WEGR) in human bladder T24 cancer cells. It was found that WEGR could inhibit the cell growth of T24 cells in a dose-dependent manner, which was associated with the induction of apoptotic cell death, as evidenced by the formation of apoptotic bodies, DNA fragmentation and increased populations of annexin-V positive cells. The induction of apoptotic cell death by WEGR was connected with an up-regulation of pro-apoptotic Bax protein expression and down-regulation of anti-apoptotic Bcl-2 and Bcl-xL proteins, and inhibition of apoptosis family proteins (XIAP, cIAP-1 and cIAP-2). In addition, apoptosis-inducing concentrations of WEGR induced the activation of caspase-9, an initiator caspase of the mitochondrial-mediated intrinsic pathway, and caspase-3, accompanied by proteolytic degradation of poly (ADP-ribose)-polymerase. WEGR also induced apoptosis via a death receptor-mediated extrinsic pathway by caspase-8 activation, resulting in the down-regulation of total Bid and suggesting the existence of cross-talk between the extrinsic and intrinsic pathways. Taken together, the present results suggest that WEGR may be a potential chemotherapeutic agent for the control of human bladder cancer cells.

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Differentiation of Dopaminergic Neurons from Mesenchymal-Like Stem Cells Derived from Human Umbilical Cord Vein

  • Kim, Ju-Ran;Lee, Jin-Ha;Jalin, Anjela Melinda;Lee, Chae-Yeon;Kang, Ah-Reum;Do, Byung-Rok;Kim, Hea-Kwon;Kam, Kyung-Yoon;Kang, Sung-Goo
    • Development and Reproduction
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    • v.13 no.3
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    • pp.173-181
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    • 2009
  • One of the most extensively studied populations of multipotent adult stem cells are mesenchymal stem cells (MSCs). MSCs derived from the human umbilical cord vein (HUC-MSCs) are morphologically and immunophenotypically similar to MSCs isolated from bone marrow. HUC-MSCs are multipotent stem cells, differ from hematopoietic stem cells and can be differentiated into neural cells. Since neural tissue has limited intrinsic capacity of repair after injury, the identification of alternate sources of neural stem cells has broad clinical potential. We isolated mesenchymal-like stem cells from the human umbilical cord vein, and studied transdifferentiation-promoting conditions in neural cells. Dopaminergic neuronal differentiation of HUC-MSCs was also studied. Neural differentiation was induced by adding bFGF, EGF, dimethyl sulfoxide (DMSO) and butylated hydroxyanisole (BHA) in N2 medium and N2 supplement. The immunoreactive cells for $\beta$-tubulin III, a neuron-specific marker, GFAP, an astrocyte marker, or Gal-C, an oligodendrocyte marker, were found. HUC-MSCs treated with bFGF, SHH and FGF8 were differentiated into dopaminergic neurons that were immunopositive for tyrosine hydroxylase (TH) antibody. HUC-MSCs treated with DMSO and BHA rapidly showed the morphology of multipolar neurons. Both immunocytochemistry and RT-PCR analysis indicated that the expression of a number of neural markers including NeuroD1, $\beta$-tubulin III, GFAP and nestin was markedly elevated during this acute differentiation. While the stem cell markers such as SCF, C-kit, and Stat-3 were not expressed after neural differentiation, we confirmed the differentiation of dopaminergic neurons by TH/$\beta$-tubulin III positive cells. In conclusion, HUC-MSCs can be differentiated into dopaminergic neurons and these findings suggest that HUC-MSCs are alternative cell source of therapeutic treatment for neurodegenerative diseases.

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Anti-Proliferative Effects of Selenium in HT-29 Colon Cancer Cells via Inhibition of Akt (HT-29 대장암세포에서 Akt 활성 저해에 따른 셀레늄의 세포 증식억제 효과)

  • Park, Song-Yi;Kim, In-Seop;Lee, Se-Hee;Lee, Sol-Hwa;Jung, Da-Woon;Park, Ock-Jin;Kim, Young-Min
    • Journal of Life Science
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    • v.22 no.1
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    • pp.55-61
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    • 2012
  • Akt is known to play an important role in cell proliferation and differentiation, and is also over-expressed in several types of cancer cells. In this study, we explored the anti-proliferative effects of selenium in HT-29 colon cancer cells, mediated through effects on Akt and COX-2. Selenium treatments at different concentrations and for different durations inhibited proliferation of HT-29 colon cancer cells and increased apoptotic cell death. Selenium treatment decreased Akt phosphorylation and COX-2 expression. Treatment with LY294002 (an Akt inhibitor) decreased proliferation of HT-29 cells, while a combined treatment with LY294002 and selenium resulted in even further decreases in cell proliferation. Inactivation of Akt by Akt siRNA treatment abolished these inhibitory effects on cell growth. COX-2 expression decreased in Akt transfected cells compared to non-transfected cells. These results suggest that selenium induced both anti-proliferative and apoptotic effects by inhibiting Akt phosphorylation and COX-2 expression. Selenium treatment also appeared to induce synergistic anti-proliferative effects by inhibition of Akt in HT-29 colon cancer cells.

Neuroprotective Effects of Acorus gramineus Soland. on Oxygen-Glucose Deprivation/Reoxygenation-Induced β-amyloid Production in SH-SY5Y Neuroblastoma Cells (허혈-재관류 유도 SH-SY5Y 모델에서 베타아밀로이드 생성에 미치는 석창포 추출물에 대한 뇌 신경보호 효과)

  • Su Young Shin;Jin-Woo Jeong;Chul Hwan Kim;Eun Jung Ahn;Seung Young Lee;Chang-Min Lee;Kyung-Min Choi
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2021.04a
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    • pp.58-58
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    • 2021
  • Although hypoxic/ischemic injury is thought to contribute to the incidence of Alzheimer disease (AD), the molecular mechanism that determines the relationship between hypoxia-induced β-amyloid (Aβ) generation and development of AD is not yet known. In this study, we investigated the protective effects of Acorus gramineus Soland. (AGS) on oxygen-glucose deprivation/reoxygenation (OGD/R)-induced A β production in SH-SY5Y human neuroblastoma cells. Pretreatment of these cells with AGS significantly attenuated OGD/R-induced production of reactive oxygen species (ROS) and elevation of levels of malondialdehyde, nitrite (NO), prostaglandin E2 (PGE2), cytokines (TNF-α, IL-1β and IL-6) and glutathione, as well as superoxide dismutase activity. AGS also reduced OGD/R-induced expression of the apoptotic protein caspase-3, the apoptosis regulator Bcl-2, and the autophagy protein becn-1. Finally, AGS reduced OGD/R-induced Aβ production and cleavage of amyloid precursor protein, by inhibiting secretase activity and suppressing the autophagic pathway. Although supporting data from in vivo studies are required, our results indicate that AGS may prevent neuronal cell damage from OGD/R-induced toxicity.

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