Induction of zinc transporters by forskolin in human trophoblast BeWo cells

The combination of sulfadiazine and pyrimethamine plus folinic acid is the conventional treatment for congenital toxoplasmosis. However, this classical treatment presents teratogenic effects and bone marrow suppression. In this sense, new therapeutic strategies are necessary to reduce these effects and improve the control of infection. In this context, biogenic silver nanoparticles (AgNp-Bio) appear as a promising alternative since they have antimicrobial, antiviral, and antiparasitic activity. The purpose of this study to investigate the action of AgNp-Bio in BeWo cells, HTR-8/SVneo cells and villous explants and its effects against Toxoplasma gondii infection. Both cells and villous explants were treated with different concentrations of AgNp-Bio or combination of sulfadiazine + pyrimethamine (SDZ + PYZ) in order to verify the viability. After, cells and villi were infected and treated with AgNp-Bio or SDZ + PYZ in different concentrations to ascertain the parasite proliferation and cytokine production profile. AgNp-Bio treatment did not reduce the cell viability and villous explants. Significant reduction was observed in parasite replication in both cells and villous explants treated with silver nanoparticles and classical treatment. The AgNp-Bio treatment increased of IL-4 and IL-10 by BeWo cells, while HTR8/SVneo cells produced macrophage migration inhibitory factor (MIF) and IL-4. In the presence of T. gondii, the treatment induced high levels of MIF production by BeWo cells and IL-6 by HTR8SV/neo. In villous explants, the AgNp-Bio treatment downregulated production of IL-4, IL-6, and IL-8 after infection. In conclusion, AgNp-Bio can decrease T. gondii infection in trophoblast cells and villous explants. Therefore, this treatment demonstrated the ability to reduce the T. gondii proliferation with induction of inflammatory mediators in the cells and independent of mediators in chorionic villus which we consider the use of AgNp-Bio promising in the treatment of toxoplasmosis in BeWo and HTR8/SVneo cell models and in chorionic villi.

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Characteristics of Calcium Uptake by BeWo Cells, a Human Trophoblast Cell Line

Placenta ◽

10.1053/plac.2001.0719 ◽

2001 ◽

Vol 22 (8-9) ◽

pp. 768-775 ◽

Cited By ~ 12

Author(s):

R. Moreau ◽

L. Simoneau ◽

J. Lafond

Keyword(s):

Cell Line ◽

Calcium Uptake ◽

Trophoblast Cell ◽

Human Trophoblast ◽

Bewo Cells ◽

Trophoblast Cell Line

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110. EFFECTS OF cAMP AND SERUM ON HUMAN TROPHOBLAST CELL VIABILITY AND DIFFERENTIATION

Reproduction Fertility and Development ◽

10.1071/srb09abs110 ◽

2009 ◽

Vol 21 (9) ◽

pp. 29

Author(s):

Y. Chen ◽

M. Allars ◽

C. Abou-Seif ◽

R. C. Nicholson

Keyword(s):

Cell Viability ◽

Syncytium Formation ◽

Culture Conditions ◽

Trophoblast Cell ◽

Bewo Cell ◽

Trophoblast Cells ◽

Human Trophoblast ◽

Serum Free ◽

Bewo Cells ◽

Free Media

The formation of syncytium is a pivotal event for trophoblast cells to interact with the placental bed. While cAMP is regarded as an inducer of syncytialisation, the affect of different culture conditions on this cAMP effect has not been explored. Therefore, the effects of cAMP on cell differentiation and viability in the presence or absence of serum were investigated in the human choriocarcinoma cell lines, BeWo and JEG-3. We observed that in the absence of cAMP, BeWo cells grew best in media containing 10% FCS, followed by media containing charcoal-stripped 10% FCS (10%CCS), and less well in serum-free media. In the presence of cAMP ( 0.25~1.5 mM ), our observations suggest different cellular programmes may be in play. Treatment of BeWo cells with 0.75 mM cAMP for 24h and 48h, in the absence of serum, increased cell viability (MTT assay) by 25.1% and 46.1% respectively, compared to the control cells. Interestingly, this cAMP effect on cell viability was not observed in the JEG-3 cell line. In contrast, BeWo cell viability was decreased by 49.5% and 25.2%, and by 27.5% and 31.1% in JEG-3 cells, when the cAMP stimulated cells were cultured for 48h in 10% CCS and 10% FCS media, respectively. In addition, we observed a change in BeWo, but not JEG-3, cell morphology to a spindle-like shape with pseudopodia when cAMP stimulated cells were cultured in media containing 10% CCS or 10% FCS for greater than 24h. Since the process of syncytialisation may involve apoptotic events, we speculate that the different effects of cAMP on cell viability in trophoblast cells may be related to syncytialising factors contained in serum media. Further study will clarify whether serum promotes syncytium formation, while the lack of serum based factors could switch the cellular programme from one of syncytialisation toward a more proliferative type.

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Comparison of folic acid uptake characteristics by human placental choriocarcinoma cells at acidic and physiological pH

Canadian Journal of Physiology and Pharmacology ◽

10.1139/y05-129 ◽

2006 ◽

Vol 84 (2) ◽

pp. 247-255 ◽

Cited By ~ 20

Author(s):

Elisa Keating ◽

Clara Lemos ◽

Isabel Azevedo ◽

Fátima Martel

Keyword(s):

Folic Acid ◽

Cytochalasin D ◽

Disulfonic Acid ◽

Acid Uptake ◽

Maternal Circulation ◽

Human Trophoblast ◽

Receptor Mediated Endocytosis ◽

Bewo Cells ◽

Carbonyl Cyanide ◽

Choriocarcinoma Cells

The aim of this work was to characterize the placental uptake of folic acid from the maternal circulation. Using 2 human trophoblast cell lines (BeWo and JAR), we verified that uptake of 3H-folic acid was pH-dependent, increasing significantly with decreasing extracellular pH. In BeWo cells, uptake of 3H-folic acid at pH 5.5 was (i) Na+-independent; (ii) inhibited by folic acid, 5-methyltetrahydrofolate (5-MTHF), and methotrexate (MTX); (iii) inhibited by the anion transport inhibitors 4,4′-diisothiocyanatostilbene-2,2′-disulphonic acid (DIDS) and 4-acetamido-4′-isothiocyano-2,2′-disulfonic acid stilbene (SITS); (iv) inhibited by the proton ionophore carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP); (v) not inhibited by blockers of receptor-mediated endocytosis (cytochalasin D and monensin); (vi) trans-inhibited by MTX and folic acid; and (vii) not affected by an anti-reduced folate transporter-1 (RFC) antibody. At pH 7.5, uptake of 3H-folic acid was (i) Na+-independent; (ii) inhibited by folic acid and MTX, but not by 5-MTHF; (iii) inhibited by SITS, but not by DIDS; (iv) not affected by FCCP; (v) inhibited by monensin (but not by cytochalasin D); (vi) trans-inhibited by folic acid (but not by MTX); and (vii) inhibited by an anti-RFC antibody. In conclusion, in BeWo cells, both RFC and receptor-mediated endocytosis seem to be involved in 3H-folic acid uptake at pH 7.5, whereas at pH 5.5, RFC and (or) a low pH-operating transporter distinct from RFC are involved.

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Cannabidiol induces apoptotic cell death in human trophoblast BeWo cells

Placenta ◽

10.1016/j.placenta.2021.07.265 ◽

2021 ◽

Vol 112 ◽

pp. e82

Author(s):

Patrícia Alves ◽

Cristina Amaral ◽

Natércia Teixeira ◽

Georgina Correia-da-Silva

Keyword(s):

Cell Death ◽

Apoptotic Cell ◽

Apoptotic Cell Death ◽

Human Trophoblast ◽

Bewo Cells

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CD9 is involved in invasion of human trophoblast-like choriocarcinoma cell line, BeWo cells

MHR Basic science of reproductive medicine ◽

10.1093/molehr/5.2.168 ◽

1999 ◽

Vol 5 (2) ◽

pp. 168-174 ◽

Cited By ~ 34

Author(s):

T. Hirano

Keyword(s):

Cell Line ◽

Human Trophoblast ◽

Bewo Cells ◽

Choriocarcinoma Cell Line

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Nobiletin, a hexamethoxyflavonoid from citrus pomace, attenuates G1 cell cycle arrest and apoptosis in hypoxia-induced human trophoblast cells of JEG-3 and BeWo via regulating the p53 signaling pathway

Food & Nutrition Research ◽

10.29219/fnr.v65.5649 ◽

2021 ◽

Author(s):

Mengling Zhang ◽

Jian Liu ◽

Rui Zhang ◽

Zengenni Liang ◽

Shenghua Ding ◽

...

Keyword(s):

Cell Cycle ◽

Western Blot ◽

Blot Analysis ◽

Cell Apoptosis ◽

Western Blot Analysis ◽

P53 Protein ◽

Human Trophoblast ◽

Protein Levels ◽

Bewo Cells ◽

Induced Apoptosis

Background: Hypoxia is associated with abnormal cell apoptosis in trophoblast cells, which causes fetal growth restriction and related placental pathologies. Few effective methods for the prevention and treatment of placenta-related diseases exist. Natural products and functional foods have always been a rich source of potential anti-apoptotic drugs. Nobiletin (NOB), a hexamethoxyflavonoid derived from the citrus pomace, shows an anti-apoptotic activity, which is a non-toxic constituent of dietary phytochemicals approved by the Food and Drug Administration. However, their effects on hypoxia-induced human trophoblast cells have not been fully studied. Objective: The aim of this study was to investigate the protective effects of NOB on hypoxia-induced apoptosis of human trophoblast JEG-3 and BeWo cells, and their underlying mechanisms. Design: First, the protective effect of NOB on hypoxia-induced apoptosis of JEG-3 and BeWo cells was studied. Cell viability and membrane integrity were determined by CCK-8 assay and lactate dehydrogenase activity, respectively. Real Time Quantitative PCR (RT-qPCR) and Western blot analysis were used to detect the mRNA and protein levels of HIF1α. Propidium iodide (PI)-labeled flow cytometry was used to detect cell cycle distribution. Cell apoptosis was detected by flow cytometry with Annexin V-FITC and PI double staining, and the expression of apoptosis marker protein cl-PARP was detected by Western blot analysis. Then, the molecular mechanism of NOB against apoptosis was investigated. Computer molecular docking and dynamics were used to simulate the interaction between NOB and p53 protein, and this interaction was verified in vitro by Ultraviolet and visible spectrum (UV-visible spectroscopy), fluorescence spectroscopy and circular dichroism. Furthermore, the changes in the expression of p53 signaling pathway genes and proteins were detected by RT-qPCR and Western blot analysis, respectively. Results: Hypoxia treatment resulted in a decreased cell viability and cell membrane integrity in JEG-3 and BeWo cell lines, and an increased expression of HIF1α, cell cycle arrest in the G1 phase, and massive cell apoptosis, which were alleviated after NOB treatment. Molecular docking and dynamics simulations found that NOB spontaneously bonded to human p53 protein, leading to the change of protein conformation. The intermolecular interaction between NOB and human p53 protein was further confirmed by UV-visible spectroscopy, fluorescence spectroscopy and circular dichroism. After the treatment of 100 μM NOB, a down-regulation of mRNA and protein levels of p53 and p21 and an up-regulation of BCL2/BAX mRNA and protein ratio were observed in JEG-3 cells; however, there was also a down-regulation of mRNA and protein levels observed for p53 and p21 in BeWo cells after the treatment of NOB. The BCL2/BAX ratio of BeWo cells did not change after the treatment of 100 μM NOB. Conclusion: NOB attenuated hypoxia-induced apoptosis in JEG-3 and BeWo cell lines and might be a potential functional ingredient to prevent pregnancy-related diseases caused by hypoxia-induced apoptosis. These findings would also suggest the exploration and utilization of citrus resources, and the development of citrus industry.

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Effect of Inflammatory Signals on Progesterone Production at the Maternal-Fetal Interface

Medical Research Archives ◽

10.18103/mra.v9i11.2584 ◽

2021 ◽

Vol 9 (11) ◽

Author(s):

R Moore ◽

D Kumar ◽

J Mansour ◽

B Mercer ◽

S Mesiano ◽

...

Keyword(s):

Structural Integrity ◽

Rupture Strength ◽

Feedback System ◽

Matrix Metalloproteinase 2 ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Human Trophoblast ◽

Gm Csf ◽

Bewo Cells ◽

Trophoblastic Cells

Introduction: In explant cultures of human fetal membranes (FM) granulocyte-macrophage-colony-stimulating-factor (GM-CSF) mediates the inflammation-induced FM weakening seen in preterm premature rupture of the membranes (pPROM) and exogenous progesterone (P4) inhibits GM-CSF and inflammation-induced FM weakening. Here we report that GM-CSF induces P4 production within the FM which then acts in a paracrine manner to counteract GM-CSF-induced weakening. Methods: FM explants mounted in Transwell inserts were cultured with control media and increasing GM-CSF, RU486 (blocks P4 action), or trilostane (blocks P4 production). P4 production, matrix metalloproteinase-2 (MMP-2) and FM rupture strength were determined. Effects of GM-CSF on P4 production and abundance of the 3ß-hydroxysteroid dehydrogenase (3ßHSD) enzyme in the BeWo human trophoblast cell line were also determined. Results: GM-CSF induced P4 production in both FM explants and BeWo cells in a concentration-dependent manner. GM-CSF also increased 3βHSD protein in BeWo cells. Incubation of FMs with RU486, or trilostane, each caused increased FM weakening. Trilostane also increased MMP-2. Exogenous P4 with trilostane repressed MMP-2 and restored FM strength. Conclusion: GM-CSF induced P4 production by FM and trophoblastic cells suggesting that locally produced P4 is increased by factors that weaken FM. Inhibition of local P4 production or action resulted in FM weakening with concomitant MMP-2 induction suggesting local P4 maintains FM structural integrity. This weakening is reversed by exogenous P4. These data are consistent with a negative-feedback system whereby P4 induced by GM-CSF, the mediator of inflammation-induced FM weakening, counteracts GM-CSF, inhibiting both its production and downstream action with resultant preservation of FM structural integrity.

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