• Title/Summary/Keyword: BeWo cells

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Activation of Phospholipase Cγ by Nitric Oxide in Choriocarcinoma Cell Line, BeWo Cells (Choriocarcinoma 세포주 BeWo 세포에서 nitric oxide에 의한 phospholipase Cγ 의 활성)

  • 차문석;곽종영
    • Journal of Life Science
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    • v.13 no.6
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    • pp.849-855
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    • 2003
  • Nitric oxide (NO) plays an important role as a signaling molecule in the proliferation of placenta trophoblasts. In this study, we investigated the effect of NO on the activation of phospholipase C (PLC) in BeWo cells, choriocar-cinoma cell line. Sodium nitroprusside (SNP), an agent to produce NO spontaneously in cells, alone increased $[^3H]$ thymidine incorporation of BeWo cells, indicating NO stimulates proliferation of the cells. NO-induced proliferation of BeWo cells was blocked by U73122, an inhibitor of PLC, suggesting that NO-induced PLC activation is involved in the cell proliferation. NO also stimulated extracellular signal-regulated kinase (ERK) in BeWo cells, indicated by increased phosphorylation of ERK1/2 in Western blotting using anti-phospho-ERK1/2 antibody. NO-induced phos-phorylation of ERK1/2 was not abrogated by U73122. $PLC\gamma_1$l but not$PLC\gamma_2$ was tyrosine phosphorylated by SNP in immunoprecipitation assay using anti-$PLC\gamma_1$/$PLC\gamma_2$ antibodies, and SNP-induced phosphorylation of $PLC\gamma_1$ was abrogated by pre-treatment of cells with genistein and PD98059, indicating that NO induced-phosphorylation of $PLC\gamma_1$ is mediated by ERK. These results suggest that NO stimulates the proliferation of BeWo cells through ERK and $PLC\gamma_1$.

Characteristics of Thiamine Uptake by the BeWo Human Trophoblast Cell Line

  • Keating, Elisa;Lemos, Clara;Azevedo, Isabel;Martel, Fatima
    • BMB Reports
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    • v.39 no.4
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    • pp.383-393
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    • 2006
  • Little is known concerning the mechanisms responsible for the transplacental transfer of thiamine. So, the aim of this work was to characterize the placental uptake of thiamine from the maternal circulation, by determining the characteristics of $^3H$-thiamine uptake by a human trophoblast cell line (BeWo). Uptake of $^3H$-thiamine (50-100 nM) by BeWo cells was: 1) temperature-dependent and energy-independent; 2) pH-dependent (uptake increased as the extracellular medium pH decreased); 3) $Na^+$-dependent and $Cl^-$-independent; 4) not inhibited by the thiamine structural analogs amprolium, oxythiamine and thiamine pyrophosphate; 5) inhibited by the unrelated organic cations guanidine, N-methylnicotinamide, tetraethylammonium, clonidine and cimetidine; 6) inhibited by the organic cation serotonin, and by two selective inhibitors of the serotonin plasmalemmal transporter (hSERT), fluoxetine and desipramine. We conclude that $^3H$-thiamine uptake by BeWo cells seems to occur through a process distinct from thiamine transporter-1 (hThTr-1) and thiamine transporter-2 (hThTr-2). Rather, it seems to involve hSERT. Moreover, chronic (48 h) exposure of cells to caffeine ($1\;{\mu}M$) stimulated and chronic exposure to xanthohumol and iso-xanthohumol (1 and $0.1\;{\mu}M$, respectively) inhibited $^3H$-thiamine uptake, these effects being not mediated through modulation of the expression levels of either hThTr-1 or hSERT mRNA.

Gas Sensing Characteristics of WO3-Doped SnO2 Thin Films Prepared by Solution Deposition Method (용액적하법으로 제조된 WO3 첨가 SnO2 박막의 가스감응 특성)

  • Choi, Joong-Ki;Cho, Pyeong-Seok;Lee, Jong-Heun
    • Korean Journal of Materials Research
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    • v.18 no.4
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    • pp.193-198
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    • 2008
  • $WO_3$-doped $SnO_2$ thin films were prepared in a solution-deposition method and their gas-sensing characteristics were investigated. The doping of $WO_3$ to $SnO_2$ increased the response ($R_a/R_g,\;R_a$: resistance in air, $R_g$: resistance in gas) to $H_2$ substantially. Moreover, the $R_a/R_g$ value of 10 ppm CO increased to 5.65, whereas that of $NO_2$ did not change by a significant amount. The enhanced response to $H_2$ and the selective detection of CO in the presence of $NO_2$ were explained in relation to the change in the surface reaction by the addition of $WO_3$. The $WO_3$-doped $SnO_2$ sensor can be used with the application of a $H_2$ sensor for vehicles that utilize fuel cells and as an air quality sensor to detect CO-containing exhaust gases emitted from gasoline engines.

Nanostructured Alloy Electrode for use in Small-Sized Direct Methanol Fuel Cells (소형 직접 메탄올 연료전지를 위한 나노 합금 전극)

  • Park Gyeong Won;Choi Jong Ho;Park In Su;Nam Woo Hyeon;Seong Yeong Eun
    • 한국전기화학회:학술대회논문집
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    • 2003.07a
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    • pp.83-88
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    • 2003
  • PtRu alloy and $PtRu-WO_3$ nanocomposite thin-film electrodes for methanol electrooxidation were fabricated by means of a sputtering method. The structural and electrochemical properties of well-defined PtRu alloy thin-film electrodes were characterized using X-ray diffraction, Rutherford backscattering spectroscopy. X-ray photoelectron spectroscopy, and electrochemical measurements. The alloy thin-film electrodes were classified as follows: Pt-based and Ru-based alloy structure. Based on structural and electrochemical understanding of the PtRu alloy thin-film electrodes, the well-controlled physical and (electro)chemical properties of $PtRu-WO_3$, showed superior specific current to that of a nanosized PtRu alloy catalyst, The homogeneous dispersion of alloy catalyst and well-formed nanophase structure would lead to an excellent catalytic electrode reaction for high-performance fuel cells. In addition, the enhanced catalytic activity in nanocomposite electrode was found to be closely related to proton transfer in tungsten oxide using in-situ electrochemical transmittance measurement.

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Anti-proliferative and angio-suppressive effect of Stoechospermum marginatum (C. Agardh) Kutzing extract using various experimental models

  • Vinayak, Rashmi;Puttananjaiah, Shilpa;Chatterji, Anil;Salimath, Bharati
    • Nutrition Research and Practice
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    • v.8 no.4
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    • pp.377-385
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    • 2014
  • BACKGROUND/OBJECTIVES: Abundant consumption of seaweeds in the diet is epidemiologically linked to the reduction in risk of developing cancer. In larger cases, however, identification of particular seaweeds that are accountable for these effects is still lacking, hindering the recognition of competent dietary-based chemo preventive approaches. The aim of this research was to establish the antiproliferative potency and angiosuppressive mode of action of Stoechospermum marginatum seaweed methanolic extract using various experimental models. MATERIALS/METHODS: Among the 15 seaweeds screened for antiproliferative activity against Ehrlich ascites tumor (EAT) cell line, Stoechospermum marginatum extract (SME) was found to be the most promising. Therefore, it was further investigated for its anti-proliferative activity in-vitro against choriocarcinoma (BeWo) and non-transformed Human embryonic kidney (HEK 293) cells, and for its anti-migratory/tube formation activity against HUVEC cells in-vitro. Subsequently, the angiosuppressive activity of S. marginatum was established by inhibition of angiogenesis in in-vivo (peritoneal angiogenesis and chorioallantoic membrane assay) and ex-vivo (rat cornea assay) models. RESULTS: Most brown seaweed extracts inhibited the proliferation of EAT cells, while green and red seaweed extracts were much less effective. According to the results, SME selectively inhibited proliferation of BeWo cells in-vitro in a dose-dependent manner, but had a lesser effect on HEK 293 cells. SME also suppressed the migration and tube formation of HUVEC cells in-vitro. In addition, SME was able to suppress VEGF-induced angiogenesis in the chorio allantoic membrane, rat cornea, and tumor induced angiogenesis in the peritoneum of EAT bearing mice. A decrease in the microvessel density count and CD31 antigen staining of treated mice peritoneum provided further evidence of its angiosuppressive activity. CONCLUSIONS: Altogether, the data underline that VEGF mediated angiogenesis is the target for the angiosuppressive action of SME and could potentially be useful in cancer prevention or treatment involving stimulated angiogenesis.

Photoelectrochamical characteristics of $WO_3$ on metal substrate for hydrogen production (텅스텐산화물/금속기판의 광전극 특성)

  • Go, GeunHo;Shinde, Pravin S.;Seo, SeonHee;Lee, Dongyoon;Lee, Wonjae
    • 한국신재생에너지학회:학술대회논문집
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    • 2011.11a
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    • pp.99.2-99.2
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    • 2011
  • Transparent conducting oxides (TCOs) supported on glass are widely used as substrates in PEC studies for photovoltaic hydrogen generation applications However, high sheet resistane ($10{\sim}15{\Omega}/cm^2$) and fragileness of glass-supported TCO substrates are the obstacles to produce the large area PEC cells. Such internal sheet resistance is detrimental to efficient collection of photogenerated majority charge carriers at the photoactive material and electrolyte interface. Moreover, these TCO substrates are very expensive and consume about 40~60% cost of the devices. Hence, a low sheet resistance of the substrate is a key point in improving the performance of PEC devices. Metallic substrates coated with a photoactive material would be a good choice for efficient charge collection. Such metal substrates based photanodes are best candidate for large-scale phtoelectrochemical water splitting for hydrogen generation. In this study, we report the enhanced PEC performance of $WO_3$ film on metal(chemical etched, bare) substrate. It is proposed that interface between $WO_3$ and the metal substrate is responsible for efficient charge transfer and demonstrated significant improvement in the photoelectrochmical performance. X-ray diffration and FESEM suduies reveled that $WO_3$ films are monoclinic, porous, polycrystalline with average grain size of ~50nm. Photocurrent of $WO_3$ prepared on metal substrates was measured in 0.5M $H_2SO_4$ electroyte under simulated $100mW/cm^2$ illumination.

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DNA Damage and Micronuclei Induced by Di (2-ethylhexyl) phthalate in Human Breast Carcinoma MCF-7 cells (Di(2-ethylhexyl) phthalate에 의해 유도된 DNA손상과 소핵 형성)

  • 김종원;한의식;박미선;엄미옥;김인숙;전혜승;정해관;심웅섭;오혜영
    • Environmental Mutagens and Carcinogens
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    • v.21 no.1
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    • pp.34-43
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    • 2001
  • Di-2-ethylhexyl phthalate (DEHP) is the most commonly used phthalate ester in polyvinyl chloride formulations including food packing and storage of human blood. DEHP is a well known as non-genotoxic carcinogen and endocrine disrupting chemical (EDC). DEHP have shown all negative results in ICH-guildeline recommended standard genotoxicity test battery. In this study, to assess the clastogenic and DNA damaging effect in human-derived tissue specific cells, DEHP was treated in human derived MCE-7 cells, HepG2 cells, LNCap cells, BeWo cells, MCE-10A cells, and female peripheral blood cells using micronucleus assay and in human breast carcinoma MCF-7 cells up to $1.28$\times$10^{-2}$ M using Comet assay. The in vitro micronucleus assay is a mutagenicity test system for the detection of chemicals which induce the formation of small membrane bound DNA fragment i.e. micronuclei in the cytoplasm of interphase cells, originated from clastogenic and/or aneugenic mechanism. The single cell gel electrophoresis assay (Comet assay) is used to detect DNA strand-breaks and alkaline labile site. In our results, DEHP increased significantly and/or dose-depentently and time-dependently micronucleus frequency at the 6 and 24 hr without metabolic activation system only in MCE-7 cells. DEHP treated with 2 hrs in MCF-7 cells using Comet assay induced DNA damage dose-depentantly.

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Differential expression of the metastasis suppressor KAI1 in decidual cells and trophoblast giant cells at the feto-maternal interface

  • Koo, Tae Bon;Han, Min-Su;Tadashi, Yamashita;Seong, Won Joon;Choi, Je-Yong
    • BMB Reports
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    • v.46 no.10
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    • pp.507-512
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    • 2013
  • Invasion of trophoblasts into maternal uterine tissue is essential for establishing mature feto-maternal circulation. The trophoblast invasion associated with placentation is similar to tumor invasion. In this study, we investigated the role of KAI1, an anti-metastasis factor, at the maternal-fetal interface during placentation. Mouse embryos were obtained from gestational days 5.5 (E5.5) to E13.5. Immunohistochemical analysis revealed that KAI1 was expressed on decidual cells around the track made when a fertilized ovum invaded the endometrium, at days E5.5 and E7.5, and on trophoblast giant cells, along the central maternal artery of the placenta at E9.5. KAI1 in trophoblast giant cells was increased at E11.5, and then decreased at E13.5. Furthermore, KAI1 was upregulated during the forskolin-mediated trophoblastic differentiation of BeWo cells. Collectively, these results indicate that KAI1 is differentially expressed in decidual cells and trophoblasts at the maternal-fetal interface, suggesting that KAI1 prevents trophoblast invasion during placentation.

Effect of the neuroprotetion and anti-Alzheimer's disease in CT99-induced PC12 cells by Gakamgobonhwan water extract (CT99 발현 PC12 세포주에서 가감고본환의 신경보호 및 항치매 효과)

  • Ahn, Dae-Gwang;Lee, So-Yeon;Yoon, Hyeon-Deok;Shin, Wo-Cheol;Park, Chang-Gook;Park, Chi-Sang
    • Herbal Formula Science
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    • v.13 no.1
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    • pp.35-48
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    • 2005
  • Alzheimer's disease(AD) is a geriatric dementia that is widespread in old age. In the near future AD will be the biggest problem in public health service. It has been widely believed that $A{\beta}$ peptide devided from APP causes apoptotic neurotoxicity in brain. However, recent evidence suggests that n99 may be an important factor causing neurotoxicity in AD. Mouse PC12 cells expressed with n99 exhibited remarkable apoptotic cell damage. We invesgated the protective effects of Gagamgobonhwan water extract(GKG). Findings from our experiments have shown that GKG inhibits the activities of CT99, which has neurotoxicities and apoptotic activities in cell line. In addition, treatment of GKG($75{\mu}g/ml$ 24 hours) partially prevented CT99-induced cytotoxicity in PC12 cells. As the result of this study, in GKG group the apoptosis in the nervous system was inhibited, the repair against the degerneration of PC12 cells by CT99 expression is promoted. Taken together, GKG exhibited inhibition of CT99-induced apoptotic cell death. GKG may be beneficial for the treatment of AD.

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Serum exosomal miR-192 serves as a potential detective biomarker for early pregnancy screening in sows

  • Ruonan Gao;Qingchun Li;Meiyu Qiu;Su Xie;Xiaomei Sun;Tao Huang
    • Animal Bioscience
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    • v.36 no.9
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    • pp.1336-1349
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    • 2023
  • Objective: The study was conducted to screen differentially expressed miRNAs in sows at early pregnancy by high-throughput sequencing and explore its mechanism of action on embryo implantation. Methods: The blood serum of pregnant and non-pregnant Landrace×Yorkshire sows were collected 14 days after artificial insemination, and exosomal miRNAs were purified for high throughput miRNA sequencing. The expression patterns of 10 differentially expressed (DE) miRNAs were validated by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The qRT-PCR quantified the abundance of serum exosomal miR-192 in pregnant and control sows, and the diagnostic power was assessed by receiver operating characteristic (ROC) analysis. The target genes of DE miRNAs were predicted with bioinformatics software, and the functional and pathway enrichment analysis was performed on gene ontology and the Kyoto encyclopedia of genes and genomes terms. Furthermore, a luciferase reporter system was used to identify the target relation between miR-192 and integrin alpha 4 (ITGA4), a gene influencing embryo implantation in pigs. Finally, the expression levels of miRNAs and the target gene ITGA4 were analyzed by qRT-PCR, and western blot, with the proliferation of BeWo cells detected by cell counting kit-8 (CCK-8). Results: A total of 221 known miRNAs were detected in the libraries of the pregnant and non-pregnant sows, of which 55 were up-regulated and 67 were down-regulated in the pregnant individuals compared with the non-pregnant controls. From these, the expression patterns of 10 DE miRNAs were validated. The qRT-PCR analysis further confirmed a significantly higher expression of miR-192 in the serum exosomes extracted from pregnant sows, when compared to controls. The ROC analysis revealed that miR-192 provided excellent diagnostic accuracy for pregnancy (area under the ROC curve [AUC]=0.843; p>0.001). The dual-luciferase reporter assay indicated that miR-192 directly targeted ITGA4. The protein expression of ITGA4 was reduced in cells that overexpressed miR-192. Overexpression of miR-192 resulted in the decreased proliferation of BeWo cells and regulated the expression of cell cycle-related genes. Conclusion: Serum exosomal miR-192 could serve as a potential biomarker for early pregnancy in pigs. miR-192 targeted ITGA4 gene directly, and miR-192 can regulate cellular proliferation.