scholarly journals Small Guanosine Triphospatase RhoA and Rho-Associated Kinase as Regulators of Trophoblast Migration

2002 ◽  
Vol 87 (12) ◽  
pp. 5808-5816 ◽  
Author(s):  
Shigetatsu Shiokawa ◽  
Mitsutoshi Iwashita ◽  
Yoshihiro Akimoto ◽  
Shinya Nagamatsu ◽  
Ken Sakai ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Stress Fibers ◽  
Extravillous Trophoblast ◽  
Dependent Manner ◽  
Focal Contact ◽  
Trophoblast Cells ◽  
Human Trophoblast ◽  
Trophoblast Migration ◽  
Human Trophoblast Cells ◽  
Actin Stress Fibers

Abstract The small guanosine triphosphatase Rho controls cell adhesion and motility through reorganization of the actin cyto-skeleton and regulation of actomyosin contractility. Among the putative target molecules of Rho, a Rho-associated coiled coil-forming protein kinase (ROCK) is thought to participate in Rho-mediated cell adhesion and motility. In the present study, we explored the expression and function of RhoA and ROCK in human trophoblast cells. The colocalization of RhoA, cytokeratin 8/18, and cytokeratin 7 in some cells located in the decidual stromal region indicated that extravillous trophoblast cells expressed RhoA. In double staining for RhoA and ROCK in human chorionic villi, RhoA staining was strongly positive in the cytoplasm of cytotrophoblasts, whereas ROCK stained in the cytoplasm of cytotrophoblasts and syncytiotrophoblasts. Both RhoA and ROCK were stained in cytoplasma of cultured human cytotrophoblast. Cultured human trophoblast cells contained actin stress fibers that were lost after treatment with C3, an exoenzyme produced by Clostridium botulinum. Y-27632, a selective ROCK inhibitor, suppressed RhoA-induced formation of actin stress fibers and formation of focal contact in trophoblast cells. The trophoblast reacquired actin stress fibers and focal contact after withdrawal of Y-27632. Cultured human cytotrophoblast cells from 7–9 wk of gestation migrated into a fibronectin-coated membrane. Both C3 exoenzyme and Y-27632 inhibited cytotrophoblast migration in a dose-dependent manner. In conclusion, cyto-trophoblasts express RhoA and ROCK in their cytoplasm, and RhoA-ROCK is involved in their assembly of actin stress fibers. Suppression of RhoA-ROCK reduces trophoblast migration. These findings suggest that RhoA-ROCK signaling is a key regulator of trophoblast cell migration.

scholarly journals Characterization of Morphological and Cytoskeletal Changes in Trophoblast Cells Induced by Insulin-Like Growth Factor-I

2002 ◽  
Vol 87 (12) ◽  
pp. 5751-5759 ◽  
Author(s):  
Maryam Kabir-Salmani ◽  
Shigetatsu Shiokawa ◽  
Yoshihiro Akimoto ◽  
Habib Hasan-Nejad ◽  
Keiji Sakai ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Rgd Peptide ◽  
Stress Fibers ◽  
Immunofluorescent Staining ◽  
Dependent Manner ◽  
Trophoblast Cells ◽  
Α Subunit ◽  
Igf I ◽  
Lamellipodia Formation ◽  
Actin Stress Fibers

Abstract IGF-I and IGF-II were appeared to play major roles in the adhesive and migratory events that are considered to be crucial in the implantation process. The purpose of this study was to determine the effects of IGF-I on trophoblast adhesion to extracellular matrix. Trophoblast cells obtained from early gestation at artificial abortion were incubated with the indicated doses of IGF-I at the indicated times. Trophoblast cells were treated with IGF-I in the presence or absence of RGD peptide and an antibody against α-subunit of IGF-I receptor (αIR3). Morphometric and morphological changes were studied using light and electron microscopy. Furthermore, vinculin, actin stress fibers, phosphorylated focal adhesion kinase (FAK), phosphotyrosine, and paxillin were immunolocalized in trophoblast cells after IGF-I treatment in the presence or absence of αIR3. Immunoprecipitation and anti-phosphotyrosine immunoblotting were carried out to detect the phosphorylated FAK and phosphorylated paxillin contents of the IGF-I-treated and untreated trophoblast cells. The results showed that IGF-I promoted trophoblast adhesion to fibronectin substrate in a time- and dose-dependent manner, and addition of RGD peptide and αIR3 monoclonal antibody abolished the effects of IGF-I in these cells. Morphological studies exhibited an increase in the lamellipodia formation upon IGF-I treatment, and confocal images of immunofluorescent staining revealed localization of phosphorylated FAK, paxillin, and vinculin at focal adhesions as well as redistribution of actin microfilaments and formation of actin stress fibers inside the cell. Western blotting, using antiphosphotyrosine demonstrated proteins with molecular masses of 125 kDa (FAK) and 68 kDa (paxillin) present in the IGF-I-treated cells, which were lacking in the control groups. In conclusion, these findings suggest that IGF-I can stimulate lamellipodia formation and promote adhesion of trophoblast cells to extracellular matrix by activating their adhesion molecules that must be activated within the implantation window.

scholarly journals α-Solanine Causes Cellular Dysfunction of Human Trophoblast Cells via Apoptosis and Autophagy

Toxins ◽  
2021 ◽  
Vol 13 (1) ◽  
pp. 67
Author(s):  
Zhilong Chen ◽  
Chen Li ◽  
Anwen Yuan ◽  
Ting Gu ◽  
Feng Zhang ◽  
...  
Keyword(s):  
Tube Formation ◽  
Extravillous Trophoblast ◽  
Autophagic Flux ◽  
Trophoblast Cells ◽  
Cellular Functions ◽  
Human Trophoblast ◽  
Antitumor Properties ◽  
Induced Apoptosis ◽  
Cellular Dysfunction ◽  
Human Trophoblast Cells

The trophoblast, an embryonic tissue, exerts a crucial role in the processes of implantation and placentation. Toxins in food can cause malfunction of trophoblasts, resulting in apoptosis, oxidative stress, and abnormal angiogenesis. α-solanine, a steroidal glycoalkaloid, has antitumor properties on several cancer cells. However, its effect on human trophoblasts has not been elucidated. In this study, human extravillous trophoblast HTR-8/SVneo cells were exposed to α-solanine. Cellular functions including proliferation, migration, invasion, tube formation, and apoptosis were assessed. To monitor autophagic flux, trophoblasts were transfected with a mCherry-GFP-LC3B vector using lentiviral transduction, and expression of autophagy-related biomarkers including Beclin 1, Atgl3, and microtubule-associated protein 1 light chain-3 (MAP1-LC3) were detected. The results show that application of 20 μM α-solanine or above inhibited the cell viability, migration, invasion, and tube formation of the human trophoblast. Cell cycle was arrested at S and G2/M phases in response to 30 μM α-solanine. α-solanine induced apoptosis of HTR-8/SVneo cells and triggered autophagy by increasing the autophagic gene expression and stimulating the formation of autophagosome and autophagic flux. In conclusion, α-solanine can impair the functions of human trophoblast cells via activation of cell apoptosis and autophagy.

scholarly journals Decorin-Mediated Inhibition of Human Trophoblast Cells Proliferation, Migration, and Invasion and Promotion of ApoptosisIn Vitro

2015 ◽  
Vol 2015 ◽  
pp. 1-10 ◽  
Author(s):  
Yanfen Zou ◽  
Xiang Yu ◽  
Jing Lu ◽  
Ziyan Jiang ◽  
Qing Zuo ◽  
...  
Keyword(s):  
Inhibitory Effect ◽  
Extravillous Trophoblast ◽  
Migration And Invasion ◽  
Trophoblast Cells ◽  
Human Trophoblast ◽  
The Matrix ◽  
Polymerase Chain ◽  
And Migration ◽  
Human Trophoblast Cells

Preeclampsia (PE) is a unique complication of pregnancy, the pathogenesis of which has been generally accepted to be associated with the dysfunctions of extravillous trophoblast (EVT) including proliferation, apoptosis, and migration and invasion. Decorin (DCN) has been proved to be a decidua-derived TGF-binding proteoglycan, which negatively regulates proliferation, migration, and invasiveness of human extravillous trophoblast cells. In this study, we identified a higher expression level of decorin in severe PE placentas by both real-time reverse transcription-polymerase chain reaction (qRT-PCR) and immunohistochemistry (IHC). And an inhibitory effect of decorin on proliferation, migration, and invasion and an enhanced effect on apoptosis in trophoblast cells HTR-8/SVneo and JEG-3 were validatedin vitro. Also the modulations of decorin on trophoblast cells’ metastasis and invasion functions were detected through regulating the matrix metalloproteinases (MMP2 and MMP9). Thus, we suggested that the contribution of decorin to the modulation of trophoblast cells might have implications for the pathogenesis of preeclampsia.

scholarly journals Regulation of the Matricellular Proteins CYR61 (CCN1) and NOV (CCN3) by Hypoxia-Inducible Factor-1α and Transforming-Growth Factor-β3 in the Human Trophoblast

Endocrinology ◽  
2010 ◽  
Vol 151 (6) ◽  
pp. 2835-2845 ◽  
Author(s):  
Nadine Wolf ◽  
Wei Yang ◽  
Caroline E. Dunk ◽  
Isabella Gashaw ◽  
Stephen J. Lye ◽  
...  
Keyword(s):  
Hypoxia Inducible Factor ◽  
Extravillous Trophoblast ◽  
Migration And Invasion ◽  
Ccn Proteins ◽  
Low Oxygen ◽  
Trophoblast Cells ◽  
Human Trophoblast ◽  
Placental Perfusion ◽  
Hypoxia Inducible Factor 1Α ◽  
Trophoblast Migration

It is known that a hypoxic environment is critical for trophoblast migration and invasion and is fundamental for appropriate placental perfusion. Because cysteine-rich 61 (CYR61, CCN1) and nephroblastoma overexpressed (NOV, CCN3) are expressed in the extravillous trophoblast and expression levels are deregulated in preeclampsia, we investigated their regulation properties in first-trimester placental explants and in JEG3 choriocarcinoma cells upon a physiological low oxygen tension of 1–3%. In placental explants, both proteins were expressed in the extravillous trophoblast cells and were increased upon hypoxia. JEG3 cells revealed a significant up-regulation of CYR61 and NOV intracellular as well as secreted protein upon hypoxic treatment accompanied by the stabilization of the hypoxia-inducible factor-1α (HIF-1α). Treatment with dimethyloxalylglycine to mimic hypoxia and silencing of HIF-1α using small interfering RNA revealed that only the increase in intracellular protein expression seems to be dependent on HIF-1α but obviously not the secretion process. Moreover, recombinant TGF-β3 was able to further enhance the amount of intracellular CCN proteins as well as secreted CYR61 levels under hypoxia. These results indicate that low oxygen levels trigger elevation of intracellular as well as secreted CYR61 and NOV protein probably in two independent pathways. Addition of recombinant CYR61 and NOV proteins increases migration as well as invasion properties of JEG3 trophoblast cells, which strengthen their role in supporting trophoblast migration invasion properties. In summary, CYR61 and NOV are regulated by HIF-1α and TGF-β3 in the trophoblast cell line JEG3, and their enhanced secretion could be implicated in appropriate placental invasion.

596: Determinants of invasion in human trophoblast cells

2007 ◽  
Vol 197 (6) ◽  
pp. S172
Author(s):  
Kathryn Drennan ◽  
Adrian Platts ◽  
Amelia Linneman ◽  
Graham Johnson ◽  
Stephen Krawetz
Keyword(s):  
Trophoblast Cells ◽  
Human Trophoblast ◽  
Human Trophoblast Cells

scholarly journals PPARα agonists clofibrate and gemfibrozil inhibit cell growth, down-regulate hCG and up-regulate progesterone secretions in immortalized human trophoblast cells

2004 ◽  
Vol 68 (2) ◽  
pp. 313-321 ◽  
Author(s):  
Fumie Hashimoto ◽  
Yoshinobu Oguchi ◽  
Mieko Morita ◽  
Kikumi Matsuoka ◽  
Satoru Takeda ◽  
...  
Keyword(s):  
Cell Growth ◽  
Trophoblast Cells ◽  
Human Trophoblast ◽  
Inhibit Cell Growth ◽  
Human Trophoblast Cells

Vascular-endothelial-cadherin modulates endothelial monolayer permeability

1999 ◽  
Vol 112 (12) ◽  
pp. 1915-1923 ◽  
Author(s):  
P.L. Hordijk ◽  
E. Anthony ◽  
F.P. Mul ◽  
R. Rientsma ◽  
L.C. Oomen ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Actin Cytoskeleton ◽  
Adhesion Molecule ◽  
Cell Adhesion Molecule ◽  
Transendothelial Migration ◽  
Stress Fibers ◽  
Vascular Endothelial ◽  
Monolayer Permeability ◽  
Actin Stress Fibers ◽  
Cell Cell

Vascular endothelial (VE)-cadherin is the endothelium-specific member of the cadherin family of homotypic cell adhesion molecules. VE-cadherin, but not the cell adhesion molecule platelet/endothelial cell adhesion molecule (PECAM-1), markedly colocalizes with actin stress fibers at cell-cell junctions between human umbilical vein endothelial cells. Inhibition of VE-cadherin-mediated, but not PECAM-1-mediated, adhesion induced reorganization of the actin cytoskeleton, loss of junctional VE-cadherin staining and loss of cell-cell adhesion. In functional assays, inhibition of VE-cadherin caused increased monolayer permeability and enhanced neutrophil transendothelial migration. In a complementary set of experiments, modulation of the actin cytoskeleton was found to strongly affect VE-cadherin distribution. Brief stimulation of the beta2-adrenergic receptor with isoproterenol induced a loss of actin stress fibers resulting in a linear, rather than ‘jagged’, VE-cadherin distribution. The concomitant, isoproterenol-induced, reduction in monolayer permeability was alleviated by a VE-cadherin-blocking antibody. Finally, cytoskeletal reorganization resulting from the inactivation of p21Rho caused a diffuse localization of VE-cadherin, which was accompanied by reduced cell-cell adhesion. Together, these data show that monolayer permeability and neutrophil transendothelial migration are modulated by VE-cadherin-mediated cell-cell adhesion, which is in turn controlled by the dynamics of the actin cytoskeleton.

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