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

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.

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In vitro study of placental trophoblast calcium uptake using JEG-3 human choriocarcinoma cells

Journal of Cell Science ◽

10.1242/jcs.98.3.333 ◽

1991 ◽

Vol 98 (3) ◽

pp. 333-342

Author(s):

R.S. Tuan ◽

C.J. Moore ◽

J.W. Brittingham ◽

J.J. Kirwin ◽

R.E. Akins ◽

...

Keyword(s):

Vitamin D3 ◽

Calcium Uptake ◽

In Vitro Model ◽

Dependent Manner ◽

Choriocarcinoma Cells ◽

Vitro Model ◽

Human Choriocarcinoma Cells ◽

Functional Components ◽

1,25 Dihydroxy Vitamin D3

During human fetal development, placental syncytiotrophoblasts actively transport calcium from the maternal to the fetal circulation. Two functional components, a cytosolic Ca2(+)-binding protein (CaBP) and a Ca2(+)-ATPase have been identified in the syncytiotrophoblasts of the chorionic villi. We report here the calcium uptake properties of a human choriocarcinoma cell line, JEG-3, which was used as an in vitro model cell system for the syncytiotrophoblasts. In culture, JEG-3 proliferated as large syncytial aggregates expressing typical syncytiotrophoblast markers. 45Ca uptake by JEG-3 was a substrate- and temperature-dependent, membrane-mediated active process that exhibited linear kinetics for up to 7 min. Both the CaBP and the Ca2(+)-ATPase were expressed by JEG-3, on the basis of biochemical, histochemical, immunochemical and or mRNA assays. Immunohistochemistry and in situ hybridization revealed that JEG-3 cells were heterogeneous with respect to the expression of the CaBP. The Ca2(+)-ATPase activity of JEG-3 was similar to the placental enzyme in terms of sensitivity to specific inhibitors, and was detected histochemically along the cell membrane. Fura-2 Ca2+ imaging revealed that calcium uptake by JEG-3 was not accompanied by a concomitant increase in cytosolic [Ca2+], suggesting a specific Ca2+ sequestration mechanism. The involvement of calciotropic hormonal regulation was evaluated by studying the response of JEG-3 to 1,25-dihydroxy vitamin D3. Calcium uptake was significantly stimulated in a dose-dependent manner by a 24-h treatment of the cells with 1,25-dihydroxy vitamin D3 (optimal dose approximately 0.5 nM); the CaBP level doubled whereas steady-state CaBP mRNA did not, suggesting that CaBP expression was regulated by 1,25-dihydroxy vitamin D3. These observations strongly suggest that the JEG-3 human choriocarcinoma cells should serve as a convenient in vitro model system for studying the cellular mechanism and regulation of transplacental calcium transport.

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Organization and chromosomal localization of the human platelet-derived endothelial cell growth factor gene

Molecular and Cellular Biology ◽

10.1128/mcb.11.4.2125-2132.1991 ◽

1991 ◽

Vol 11 (4) ◽

pp. 2125-2132

Author(s):

K Hagiwara ◽

G Stenman ◽

H Honda ◽

P Sahlin ◽

A Andersson ◽

...

Keyword(s):

Endothelial Cell ◽

Growth Factor ◽

Cell Growth ◽

Human Platelet ◽

Angiogenic Factor ◽

Transcription Start Sites ◽

Endothelial Cell Growth Factor ◽

Endothelial Cell Growth

Human platelet-derived endothelial cell growth factor (hPD-ECGF) is a novel angiogenic factor which stimulates endothelial cell growth in vitro and promotes angiogenesis in vivo. We report here the cloning and sequencing of the gene for hPD-ECGF and its flanking regions. This gene is composed of 10 exons dispersed over a 4.3-kb region. Its promoter lacks a TATA box and a CCAAT box, structures characteristic of eukaryotic promoters. Instead, six copies of potential Sp1-binding sites (GGGCGG or CCGCCC) were clustered just upstream of the transcription start sites. Southern blot analysis using genomic DNAs from several vertebrates suggested that the gene for PD-ECGF is conserved phylogenetically among vertebrates. The gene for hPD-ECGF was localized to chromosome 22 by analysis of a panel of human-rodent somatic cell hybrid lines.

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DARC Regulates Angiogenesis by Mediating Vascular Endothelial Cell Migration

10.21203/rs.3.rs-115340/v1 ◽

2020 ◽

Author(s):

Xiaolin Wang ◽

Yongqian Bian ◽

Yuejun Li ◽

Jing Li ◽

Congying Zhao ◽

...

Keyword(s):

Cell Migration ◽

Endothelial Cell ◽

Vascular Endothelial Cell ◽

Tube Formation ◽

Endothelial Cell Migration ◽

Immunofluorescent Staining ◽

Control Group ◽

Rhoa Activation ◽

And Control

Abstract Background: DARC (The Duffy antigen receptor for chemokines) is a kind of glycosylated membrane protein that binds to members of the CXC chemokine family associated with angiogenesis and has recently been reported to be implicated in diverse normal physiologic processes. This study aimed to investigate the involvement of DARC in angiogenesis, which is known to generate new capillary blood vessels from preexisting ones. Methods: HDMECs (Human dermal microvascular endothelial cells) were divided into two groups (DARC overexpression group, and control group). We used Brdu staining to detect cell proliferation, and wound healing assay to detect cell migration. Then tube formation assay were observed. Also, western blot and immunofluorescent staining were used to estimate the relationship between DARC and RhoA (Ras homolog gene family, member A). Results: HDMECs proliferation, migration, and tube formation were inhibited significantly when DARC was overexpressed intracellular. DARC impaired microfilament dynamics and intercellular connection in migrating cells, and RhoA activation underlay the effect of DARC on endothelial cell. Furthermore, DARC inhibited the formation of new capillaries in vitro. Conclusion: Our ﬁndings revealed the role of DARC in the angiogenic process and provided a novel mechanism for RhoA activation during endothelial cell migration and angiogenesis.

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Expression of Serine Proteinase Inhibitor PP5/TFPI-2/MSPI Decreases the Invasive Potential of Human Choriocarcinoma Cells in Vitro and in Vivo

Gynecologic Oncology ◽

10.1006/gyno.2001.6394 ◽

2001 ◽

Vol 83 (2) ◽

pp. 325-333 ◽

Cited By ~ 21

Author(s):

Ming-shou Jin ◽

Kaori Udagawa ◽

Etsuko Miyagi ◽

Tsuneo Nakazawa ◽

Fumiki Hirahara ◽

...

Keyword(s):

Proteinase Inhibitor ◽

Serine Proteinase ◽

Serine Proteinase Inhibitor ◽

Invasive Potential ◽

Choriocarcinoma Cells ◽

Human Choriocarcinoma Cells

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Intrauterine growth restriction decreases pulmonary alveolar and vessel growth and causes pulmonary artery endothelial cell dysfunction in vitro in fetal sheep

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00197.2011 ◽

2011 ◽

Vol 301 (6) ◽

pp. L860-L871 ◽

Cited By ~ 120

Author(s):

Paul J. Rozance ◽

Gregory J. Seedorf ◽

Alicia Brown ◽

Gates Roe ◽

Meghan C. O'Meara ◽

...

Keyword(s):

Endothelial Cell ◽

Cell Growth ◽

Pulmonary Artery ◽

Intrauterine Growth Restriction ◽

Tube Formation ◽

No Production ◽

Vascular Growth ◽

Intrauterine Growth ◽

Pulmonary Artery Endothelial Cell

Intrauterine growth restriction (IUGR) increases the risk for bronchopulmonary dysplasia (BPD). Abnormal lung structure has been noted in animal models of IUGR, but whether IUGR adversely impacts fetal pulmonary vascular development and pulmonary artery endothelial cell (PAEC) function is unknown. We hypothesized that IUGR would decrease fetal pulmonary alveolarization, vascular growth, and in vitro PAEC function. Studies were performed in an established model of severe placental insufficiency and IUGR induced by exposing pregnant sheep to elevated temperatures. Alveolarization, quantified by radial alveolar counts, was decreased 20% ( P < 0.005) in IUGR fetuses. Pulmonary vessel density was decreased 44% ( P < 0.01) in IUGR fetuses. In vitro, insulin increased control PAEC migration, tube formation, and nitric oxide (NO) production. This response was absent in IUGR PAECs. VEGFA stimulated tube formation, and NO production also was absent. In control PAECs, insulin increased cell growth by 68% ( P < 0.0001). Cell growth was reduced in IUGR PAECs by 29% at baseline ( P < 0.01), and the response to insulin was attenuated ( P < 0.005). Despite increased basal and insulin-stimulated Akt phosphorylation in IUGR PAECs, endothelial NO synthase (eNOS) protein expression as well as basal and insulin-stimulated eNOS phosphorylation were decreased in IUGR PAECs. Both VEGFA and VEGFR2 also were decreased in IUGR PAECs. We conclude that fetuses with IUGR are characterized by decreased alveolar and vascular growth and PAEC dysfunction in vitro. This may contribute to the increased risk for adverse respiratory outcomes and BPD in infants with IUGR.

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CYLD regulates angiogenesis by mediating vascular endothelial cell migration

Blood ◽

10.1182/blood-2009-10-248526 ◽

2010 ◽

Vol 115 (20) ◽

pp. 4130-4137 ◽

Cited By ~ 52

Author(s):

Jinmin Gao ◽

Lei Sun ◽

Lihong Huo ◽

Min Liu ◽

Dengwen Li ◽

...

Keyword(s):

Endothelial Cells ◽

Cell Migration ◽

Endothelial Cell ◽

Vascular Endothelial Cells ◽

Vascular Endothelial Cell ◽

Tube Formation ◽

Endothelial Cell Migration ◽

Vascular Endothelial

Cylindromatosis (CYLD) is a deubiquitinase that was initially identified as a tumor suppressor and has recently been implicated in diverse normal physiologic processes. In this study, we have investigated the involvement of CYLD in angiogenesis, the formation of new blood vessels from preexisting ones. We find that knockdown of CYLD expression significantly impairs angiogenesis in vitro in both matrigel-based tube formation assay and collagen-based 3-dimensional capillary sprouting assay. Disruption of CYLD also remarkably inhibits angiogenic response in vivo, as evidenced by diminished blood vessel growth into the angioreactors implanted in mice. Mechanistic studies show that CYLD regulates angiogenesis by mediating the spreading and migration of vascular endothelial cells. Silencing of CYLD dramatically decreases microtubule dynamics in endothelial cells and inhibits endothelial cell migration by blocking the polarization process. Furthermore, we identify Rac1 activation as an important factor contributing to the action of CYLD in regulating endothelial cell migration and angiogenesis. Our findings thus uncover a previously unrecognized role for CYLD in the angiogenic process and provide a novel mechanism for Rac1 activation during endothelial cell migration and angiogenesis.

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Roundabout4 Suppresses Glioma-Induced Endothelial Cell Proliferation, Migration and Tube Formation in Vitro by Inhibiting VEGR2-Mediated PI3K/AKT and FAK Signaling Pathways

Cellular Physiology and Biochemistry ◽

10.1159/000373982 ◽

2015 ◽

Vol 35 (5) ◽

pp. 1689-1705 ◽

Cited By ~ 27

Author(s):

Heng Cai ◽

Yixue Xue ◽

Zhen Li ◽

Yi Hu ◽

Zhenhua Wang ◽

...

Keyword(s):

Cell Proliferation ◽

Endothelial Cells ◽

Endothelial Cell ◽

Signaling Pathways ◽

Conditioned Medium ◽

Tube Formation ◽

Endothelial Cell Proliferation ◽

Akt Inhibitor ◽

Over Expression

Background and Aims: Endothelial cell (EC) proliferation, migration, and tube formation are the critical steps for tumor angiogenesis, which is involved in the formation of new tumor blood vessels. Roundabout4 (Robo4), a new member of Robo proteins family, is specifically expressed in endothelial cells. This study aimed to investigate the effects of Robo4 on glioma-induced endothelial cell proliferation, migration and tube formation in vitro. Methods and Results: We found that Robo4 was endogenously expressed in Human Brain Microvascular Endothelial Cells (HBMECs), while Robo4 was significantly down-regulated in endothelial cells cultured in glioma conditioned medium. Robo4 over-expression remarkably suppressed glioma-induced endothelial cell proliferation, migration and tube formation in vitro. In addition, Robo4 influenced the glioma-induced angiogenesis via binding to its ligand Slit2. Further studies demonstrated that the knockdown of Robo4 up-regulated the phosphorylation of VEGFR2, PI3K, AKT and FAK in EC cultured in glioma conditioned medium. VEGFR2 inhibitor SU-1498, AKT inhibitor LY294002 and FAK inhibitor 14 (FAK inhibitor) blocked the Robo4 knockdown-mediated alteration in glioma angiogenesis in vitro. Conclusion: Our results proved that Robo4 suppressed glioma-induced endothelial cell proliferation, migration and tube formation in vitro by inhibiting VEGR2-mediated activation of PI3K/AKT and FAK signaling pathways.

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In vitro angiogenesis: endothelial cell tube formation on gelled basement membrane extract

Nature Protocols ◽

10.1038/nprot.2010.6 ◽

2010 ◽

Vol 5 (4) ◽

pp. 628-635 ◽

Cited By ~ 377

Author(s):

Irina Arnaoutova ◽

Hynda K Kleinman

Keyword(s):

Endothelial Cell ◽

Basement Membrane ◽

Tube Formation ◽

Basement Membrane Extract ◽

Membrane Extract ◽

In Vitro Angiogenesis

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Monocyte Derivatives Promote Angiogenesis and Myocyte Survival in a Model of Myocardial Infarction

Cell Transplantation ◽

10.3727/096368909x484266 ◽

2010 ◽

Vol 19 (4) ◽

pp. 369-386 ◽

Cited By ~ 24

Author(s):

M. Bouchentouf ◽

P. Paradis ◽

K. A. Forner ◽

J. Cuerquis ◽

M. N. Boivin ◽

...

Keyword(s):

Myocardial Infarction ◽

Endothelial Cell ◽

Mononuclear Cells ◽

Endothelial Cell Proliferation ◽

Blood Monocytes ◽

Vascular Networks ◽

Peripheral Blood Mononuclear ◽

Mature Endothelial Cell ◽

Tnf Α

In this study, we have investigated the hypothesis that previously reported beneficial effect of peripheral blood mononuclear cells cultured under angiogenic conditions on cardiovascular function following ischemia is not limited to EPCs but also to monocytes contained therein. We first purified and analyzed the phenotype and secretome of human and murine blood monocytes cultured under angiogenic conditions (named MDs for monocyte derivatives) and tested their effect in a mouse model of myocardial infarction (MI). FACS analysis of MDs shows that these cells express mature endothelial cell markers and that their proliferative capacity is virtually absent, consistent with their end-differentiated monocytic ontogeny. MDs secreted significant levels of HGF, IGF-1, MCP-1, and sTNFR-1 relative to their monocyte precursors. MDs were unable to form vascular networks in vitro when cultured on matrix coated flasks. Treatment of murine HL-1 cardiomyocyte cell line with MD-conditioned medium reduced their death induced by TNF-α, staurosporine, and oxidative stress, and this effect was dependent upon MD-derived sTNFR-1, HGF, and IGF-1. We further demonstrate that MD secretome promoted endothelial cell proliferation and capacity to form vessels in vitro and this was dependent upon MD-derived MCP-1, HGF, and IGF-1. Echocardiography analysis showed that MD myocardial implantation improved left ventricle fractional shortening of mouse hearts following MI and was associated with reduced myocardial fibrosis and enhancement of angiogenesis. Transplanted MDs and their secretome participate in preserving functional myocardium after ischemic insult and attenuate pathological remodeling.

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