Characterization of the human c-fms gene product and its expression in cells of the monocyte-macrophage lineage

1985 ◽  
Vol 5 (12) ◽  
pp. 3458-3466
Author(s):  
J Woolford ◽  
V Rothwell ◽  
L Rohrschneider
Keyword(s):  
Rabbit Antiserum ◽  
Growth Factor Receptor ◽  
Protein Product ◽  
Human Cells ◽  
Cell Protein ◽  
Bewo Cell ◽  
Metabolic Labeling ◽  
Bewo Cells ◽  
Human Choriocarcinoma Cells

The McDonough strain of feline sarcoma virus contains an oncogene called v-fms whose ultimate protein product (gp140v-fms) resembles a cell surface growth factor receptor. To identify and characterize the protein product of the proto-oncogene c-fms, antisera were prepared to the viral fms sequences and used to detect specific cross-reacting sequences in human choriocarcinoma cells (BeWo) known to express c-fms mRNA. Both tumor-bearing rat sera and a rabbit antiserum prepared to a segment of v-fms expressed in Escherichia coli detected a 140-kilodalton (kDa) glycoprotein in the BeWo cells. Tryptic fingerprint analysis of [35S]methionine-labeled proteins indicated that the viral fms proteins and the 140-kDa BeWo cell protein were highly related. This 140-kDa glycoprotein contained an associated tyrosine kinase activity in vitro and was labeled principally on serine after 32Pi metabolic labeling. These results suggest that the 140-kDa protein in BeWo cells is the protein product of the human c-fms proto-oncogene. This conclusion is supported by the finding that a similar protein is detectable only in other human cells that express c-fms mRNA. These other human cells include adherent monocytes and the cell line ML-1, which can be induced to differentiate along the monocyte-macrophage pathway. This is in agreement with current thought that the c-fms proto-oncogene product functions as the CSF-1 receptor specific to this pathway.

scholarly journals Characterization of the human c-fms gene product and its expression in cells of the monocyte-macrophage lineage.

1985 ◽  
Vol 5 (12) ◽  
pp. 3458-3466 ◽  
Author(s):  
J Woolford ◽  
V Rothwell ◽  
L Rohrschneider
Keyword(s):  
Rabbit Antiserum ◽  
Growth Factor Receptor ◽  
Protein Product ◽  
Human Cells ◽  
Cell Protein ◽  
Bewo Cell ◽  
Metabolic Labeling ◽  
Bewo Cells ◽  
Human Choriocarcinoma Cells

The McDonough strain of feline sarcoma virus contains an oncogene called v-fms whose ultimate protein product (gp140v-fms) resembles a cell surface growth factor receptor. To identify and characterize the protein product of the proto-oncogene c-fms, antisera were prepared to the viral fms sequences and used to detect specific cross-reacting sequences in human choriocarcinoma cells (BeWo) known to express c-fms mRNA. Both tumor-bearing rat sera and a rabbit antiserum prepared to a segment of v-fms expressed in Escherichia coli detected a 140-kilodalton (kDa) glycoprotein in the BeWo cells. Tryptic fingerprint analysis of [35S]methionine-labeled proteins indicated that the viral fms proteins and the 140-kDa BeWo cell protein were highly related. This 140-kDa glycoprotein contained an associated tyrosine kinase activity in vitro and was labeled principally on serine after 32Pi metabolic labeling. These results suggest that the 140-kDa protein in BeWo cells is the protein product of the human c-fms proto-oncogene. This conclusion is supported by the finding that a similar protein is detectable only in other human cells that express c-fms mRNA. These other human cells include adherent monocytes and the cell line ML-1, which can be induced to differentiate along the monocyte-macrophage pathway. This is in agreement with current thought that the c-fms proto-oncogene product functions as the CSF-1 receptor specific to this pathway.

Ultrastructure of Choriocarcinoma in Vitro

1969 ◽  
Vol 27 ◽  
pp. 362-363
Author(s):  
John C. Garancis ◽  
R. A. Pattillo
Keyword(s):  
Cell Line ◽  
Physiological Function ◽  
Cell System ◽  
Bewo Cell ◽  
Bewo Cells ◽  
Gestational Choriocarcinoma ◽  
Bewo Cell Line

Growth of cell system (BeWo-cell line) derived from human gestational choriocarcinoma has been established and continuously maintained in-vitro. Furthermore, it is evident from the previous studies that this cell line has retained the physiological function of the placental trophoblasts, namely the synthesis of human chorionic gonadotrophil(HCG).The BeWo cells were relatively small and possessed single nuclei, thus indicating that this cell line consists exclusively of cytotrophoblasts. In some instances cells appeared widely separated and their lateral surfaces were provided with numerous microvilli (Fig.1).

scholarly journals Resistin regulates human choriocarcinoma cell invasive behaviour and endothelial cell angiogenic processes

2006 ◽  
Vol 189 (3) ◽  
pp. 691-699 ◽  
Author(s):  
Nicoletta Di Simone ◽  
Fiorella Di Nicuolo ◽  
Maurizio Sanguinetti ◽  
Roberta Castellani ◽  
Marco D’Asta ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Mononuclear Cells ◽  
Tube Formation ◽  
Angiogenic Factor ◽  
Tissue Inhibitors Of Metalloproteinases ◽  
Bewo Cell ◽  
Choriocarcinoma Cells ◽  
Cell Invasiveness ◽  
Human Choriocarcinoma Cells

Resistin is a novel hormone that is secreted by human adipocytes and mononuclear cells and is probably associated with insulin resistance. Recently, resistin has been postulated to play a role in pregnancy, and resistin gene expression has been observed in placental tissues. However, it is still not known if resistin is able to affect trophoblast functions and development. Therefore, we investigated the hypothesis that resistin might regulate trophoblast production of matrix metalloproteinases (MMPs), the tissue inhibitors of metalloproteinases (TIMPs), trophoblast invasive behavior and the angiogenic processes. In human choriocarcinoma cells (BeWo), resistin (10–100 ng/ml) enhanced both MMP-2 protein and mRNA expression, significantly reduced TIMP-1 and TIMP-2 and increased trophoblast-like cell invasiveness. We analyzed the effect of resistin on an in vitro angiogenesis system for endothelial cells (HUVEC) and we evaluated its ability to modulate the secretion of an angiogenic factor, vascular endothelial growth factor (VEGF). Our data showed that resistin induced VEGF production and we observed that the addition of resistin stimulated endothelial cell tube formation. These findings suggest that resistin might be able to induce BeWo cell invasiveness and to contribute to the control of placental vascular development.

scholarly journals Fusarium Mycotoxins Disrupt the Barrier and Induce IL-6 Release in a Human Placental Epithelium Cell Line

Toxins ◽  
2019 ◽  
Vol 11 (11) ◽  
pp. 665 ◽  
Author(s):  
Negisa Seyed Toutounchi ◽  
Astrid Hogenkamp ◽  
Soheil Varasteh ◽  
Belinda van’t Land ◽  
Johan Garssen ◽  
...  
Keyword(s):  
Bewo Cell ◽  
Placental Barrier ◽  
Fusarium Mycotoxins ◽  
Barrier Integrity ◽  
Bewo Cells ◽  
Junctional Proteins ◽  
Proinflammatory Responses ◽  
Vitro Model ◽  
Cytotoxicity Effects

Deoxynivalenol, T-2 toxin, and zearalenone, major Fusarium mycotoxins, contaminate human food on a global level. Exposure to these mycotoxins during pregnancy can lead to abnormalities in neonatal development. Therefore, the aim of this study was to investigate the effects of Fusarium mycotoxins on human placental epithelial cells. As an in vitro model of placental barrier, BeWo cells were exposed to different concentrations of deoxynivalenol, zearalenone or T-2 toxin. Cytotoxicity, effects on barrier integrity, paracellular permeability along with mRNA and protein expression and localization of junctional proteins after exposure were evaluated. Induction of proinflammatory responses was determined by measuring cytokine production. Increasing mycotoxin concentrations affect BeWo cell viability, and T-2 toxin was more toxic compared to other mycotoxins. Deoxynivalenol and T-2 toxin caused significant barrier disruption, altered protein and mRNA expression of junctional proteins, and induced irregular cellular distribution. Although the effects of zearalenone on barrier integrity were less prominent, all tested mycotoxins were able to induce inflammation as measured by IL-6 release. Overall, Fusarium mycotoxins disrupt the barrier of BeWo cells by altering the expression and structure of junctional proteins and trigger proinflammatory responses. These changes in placental barrier may disturb the maternal–fetal interaction and adversely affect fetal development.

scholarly journals ZBED1 Regulates Genes Important for Multiple Biological Processes of the Placenta

Genes ◽  
2022 ◽  
Vol 13 (1) ◽  
pp. 133
Author(s):  
Simone Johansen ◽  
Sofie Traynor ◽  
Malene Laage Ebstrup ◽  
Mikkel Green Terp ◽  
Christina Bøg Pedersen ◽  
...  
Keyword(s):  
Cell Model ◽  
Trophoblast Cell ◽  
Innate Immune ◽  
Bewo Cell ◽  
Biological Processes ◽  
Bewo Cells ◽  
Genes Encoding ◽  
And Function

The transcription factor ZBED1 is highly expressed in trophoblast cells, but its functions in the processes of trophoblast and placental biology remain elusive. Here, we characterized the role of ZBED1 in trophoblast cell differentiation using an in vitro BeWo cell model. We demonstrate that ZBED1 is enhanced in its expression early after forskolin-induced differentiation of BeWo cells and regulates many of the genes that are differentially expressed as an effect of forskolin treatment. Specifically, genes encoding markers for the differentiation of cytotrophoblast into syncytiotrophoblast and factors essential for trophoblast cell fusion and invasion were negatively regulated by ZBED1, indicating that ZBED1 might be important for maintaining a steady pool of cytotrophoblast cells. In addition, ZBED1 affected genes involved in the regulation of trophoblast cell survival and apoptosis, in agreement with the observed increase in apoptosis upon knockdown of ZBED1 in forskolin-treated BeWo cells. In addition, genes implicated in the differentiation, recruitment, and function of innate immune cells by the placenta were affected by ZBED1, further suggesting a role for this protein in the regulation of maternal immune tolerance. In conclusion, our study implicates ZBED1 in major biological processes of placental biology.

217 THE EXPRESSION OF β-OXIDATION RELATING GENES UNDER HYPOXIC STRESS IN HUMAN PLACENTAL CHORIOCARCINOMA CELL (BeWo)

2015 ◽  
Vol 27 (1) ◽  
pp. 198
Author(s):  
E.-K. Shin ◽  
E.-B. Jeung
Keyword(s):  
Hypoxic Condition ◽  
Bewo Cell ◽  
Hypoxic Stress ◽  
Experimental Conditions ◽  
Fluorescent Intensity ◽  
Peripheral Tissues ◽  
Bewo Cells ◽  
Normoxic Control ◽  
La Jolla

Preeclampsia (PE) is thought in many cases to be caused by a shallowly implanted placenta that becomes hypoxic. Hypoxia can result from a failure at any stage in the delivery of oxygen to the cells. In peripheral tissues, oxygen diffuses down a pressure gradient into cells and moves into their mitochondria, where it is used to produce energy in conjunction with the breakdown of glucose, fats, and some amino acids. The aim of the study was to investigate the question of whether hypoxic stress is involved in β-oxidation of human placental BeWo cells. One of the β-oxidation related genes, ACADVL was detected by gene-fishing technology using the preeclamptic placenta of human. We conducted in vitro experiments to confirm a preliminary study by inducing hypoxic stress in the human placental BeWo cell. BeWo cells were cultured at 37°C in a 20% O2, 5% CO2 humidified tissue culture incubator. And then we induced hypoxic stress in BeWo cell cultured under 1% O2, 5% CO2, and balanced with N2. The BeWo cells were divided into three groups: normoxia, hypoxia 24 h, and hypoxia 48 h. The expression of β-oxidation related genes (ACADVL, EHHADH, HADH, ACAA) were observed under hypoxic condition in BeWo cells by using real-time RT-PCR. Relative quantification with HPRT1 was based on the comparison of CT at a constant fluorescent intensity. The amount of transcript is inversely related to the observed CT, and for every 2-fold dilution in the transcript, CT is expected to increase by 1. Relative expression was calculated using the equation R = 2–(CTsample – CTcontrol). Data were analysed with a nonparametric one-way ANOVA, followed by Tukey's test for multiple comparisons. All statistical analyses were performed using GraphPad Prism software (v 4.0, GraphPad Software, La Jolla, CA, USA). P-values <0.05 were considered statistically significant. The expression of a gene known as a biomarker for hypoxia, HIF-1a, was significantly increased in BeWo cells which induced preeclampsia. The elevated level of HIF-1a indicates that our experimental conditions closely mimicked preeclampsia. The β-oxidation related genes, ACADVL, EHHADH, and HADH expressions were significantly increased under hypoxic condition in BeWo compared with normoxic control. These results indicate that changes of β-oxidation related genes observed under hypoxic BeWo cells are similar to ones associated with preeclampsia, and the expression of β-oxidation related genes were up-regulated by hypoxic stress. They might be involved in pathogenesis of preeclampsia during gestation. Taken together, increase of β-oxidation-related genes under hypoxic stress may cause a compensation of energy metabolism deficiency through lipid metabolism.

scholarly journals The UL4 gene of pseudorabies virus encodes a minor infected-cell protein that is dispensable for virus replication

2006 ◽  
Vol 87 (9) ◽  
pp. 2517-2525 ◽  
Author(s):  
Walter Fuchs ◽  
Harald Granzow ◽  
Robert Klopfleisch ◽  
Barbara G. Klupp ◽  
Thomas C. Mettenleiter
Keyword(s):  
Pseudorabies Virus ◽  
Rabbit Antiserum ◽  
Human Herpesvirus ◽  
Rabbit Kidney ◽  
Cell Protein ◽  
Growth Defect ◽  
Kidney Cells ◽  
Structural Protein ◽  
Herpesvirus 1

Although homologues of the open reading frame (ORF) UL4 of herpes simplex virus 1 (Human herpesvirus 1) have been found in the genomes of all hitherto-analysed alphaherpesviruses, little is known about their function. In a project to analyse systematically, in an isogenic and standardized assay system, the gene products of the alphaherpesvirus pseudorabies virus (PrV; Suid herpesvirus 1), the PrV UL4 gene product was identified using a monospecific rabbit antiserum prepared against a bacterial fusion protein. Western blot and immunofluorescence analyses revealed that the 146 codon UL4 ORF of PrV was translated into a nuclear 15 kDa protein which was detectable from 6 h after infection of rabbit kidney cells, but was not found in purified virus particles. For functional analysis, a UL4-negative virus recombinant (PrV-ΔUL4F) was generated by mutagenesis of an infectious full-length clone of the PrV genome in E. coli. PrV-ΔUL4F was replication-competent in rabbit kidney cells, and plaque formation was not affected by the mutation. However, maximum virus titres of PrV-ΔUL4F were decreased about fivefold compared with wild-type PrV, and electron microscopy of infected cells demonstrated an impairment of release of mature virions. This growth defect of PrV-ΔUL4F could be corrected completely by propagation in UL4-expressing cells. Correlating with the inconspicuous in vitro phenotype, neurovirulence of PrV-ΔUL4F was also not affected significantly. Thus, UL4 encodes a non-structural protein of PrV that enhances virion formation but is not essential for PrV replication in vitro or in vivo.

scholarly journals The use of BeWo cells as an in vitro model for placental iron transport

2008 ◽  
Vol 295 (5) ◽  
pp. C1445-C1453 ◽  
Author(s):  
Sarah J. Heaton ◽  
John J. Eady ◽  
Mary L. Parker ◽  
Kathryn L. Gotts ◽  
Jack R. Dainty ◽  
...  
Keyword(s):  
Total Dose ◽  
Iron Transport ◽  
In Vitro Model ◽  
Cell Layer ◽  
Bewo Cell ◽  
Apical Surface ◽  
Bewo Cells ◽  
Cell Layers ◽  
Vitro Model

BeWo cells are a placental cell line that has been widely used as an in vitro model for the placenta. The b30 subclone of these cells can be grown on permeable membranes in bicameral chambers to form confluent cell layers, enabling rates of both nutrient uptake into the cells from the apical surface and efflux from the basolateral membrane to be determined. The aim of this study was to evaluate structural and functional properties of confluent b30 BeWo cell layers grown in bicameral chambers, focusing on the potential application for studying receptor-mediated uptake and transport of transferrin (Tf)-bound iron (Fe-Tf). While it proved extremely difficult to establish and maintain an intact BeWo cell monolayer, it was possible to grow the cells to a confluent multilayer. Iron, applied as Fe-Tf, was rapidly transported across this cell layer; 9.3 ± 0.5% of the total dose was transported after 8 h, equivalent to 38.8 ± 2.1 pmol·cm−2·h−1. Transfer of Tf across the cell layer was much more limited; 2.4 ± 0.2% of the total dose was transported after 8 h, equivalent to 5.0 ± 0.4 pmol·cm−2·h−1. Compartmental modeling of these data suggested that iron was transported across the cell layer predominantly, if not exclusively, via a transcellular route, whereas Tf taken up into the cells was predominantly recycled back to the apical compartment. The results suggest that these cells are very efficient at transporting iron and, under carefully controlled conditions, can be a valuable tool for the study of iron transport in the placenta.

Sign in / Sign up

Export Citation Format

Share Document