Steroid regulation of oncofetal fibronectin expression in human cytotrophoblasts

Guller, Seth; LaCroix, Noelle C.; Kirkun, Graciela; Wozniak, Robert; Markiewicz, Leszek; Wang, En-Yu; Kaplan, Paul; Lockwood, Charles J.

doi:10.1016/0960-0760(93)90202-8

Cited by 44 publications

(43 citation statements)

References 26 publications

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“…The concentrations of DEX used have already been shown to affect synthesis of extracellular matrix proteins in terms of trophoblast and gal-1 in HTR-8/SVneo cells (11,25,26). The data obtained over a wide range of concentrations (0.1-100 nmol/L) in this study showed that DEX had a stimu latory effect on proliferation in both cell types.…”

Section: Discussionsupporting

confidence: 49%

Choriocarcinoma cell line Response to Dexamethasone

Bojić‐Trbojević¹,

Kolundžić²,

Petronijević³

et al. 2010

Journal of Medical Biochemistry

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Kratak sadr`aj: Horiokarcinomske }elijske linije JAr i JEG-3 su model sistemi za ispitivanje transformisanog trofo blasta. Obe }elijske linije eksprimiraju galektin-1, ~ija je pove}ana ekspresija pokazana u horiokarcinomima u odno su na normalni trofoblast trudno}e. U ovom radu ispitivan je efekat sintetskog glukokortikoida deksametazona na JAr i JEG-3 }elijske linije, upotrebom MTT testa, ELISA testa na }elijama i testovima adhezije i migracije. Broj `ivih }elija, kao indikator proliferacije }elija, zna ~ajno je pove}an nakon tretmana 0,1 i 1 nmol/L deksameta zonom, a proliferacija JEG-3 je zna~ajno pove}ana u ~itavom opsegu upotrebljenih koncentracija deksameta zo na (0,1-100 nmol/L). Galektin-1 je u JAr moduliran deksametazonom, koji blago, ali zna~ajno smanjuje pro duk ciju galektina-1 u niskim koncentracijama (0,1 i 1 nmol/L). Kod JEG-3 }elija, produkcija galektina-1 je zna~ajno smanjena samo nakon tretmana 100 nmol/L deksameta zonom. U prisustvu laktoze kao inhibitornog {e}era za galektin-1 adhezija JEG-3 }elija je zna~ajno pove}ana, dok deksa metazon uz to smanjuje i migraciju JEG-3 }elija. Rezultati dobijeni u ovom radu po kazuju da deksametazon uti~e na proliferaciju i u manjoj meri na galektin-1 kod JAr i JEG-3 }elija, na na~in spe ci fi~an za }elijsku liniju. Dobijeni podaci ukazuju na to da bi tret man glukokortikoidima in vivo mogao imati uticaja na }elije horiokarcinoma. Summary: Choriocarcinoma cell lines JAr and JEG-3 are model systems for the study of transformed trophoblast. Both cell lines were shown to produce galectin-1, expression of which was increased in choriocarcinoma when compared to the normal trophoblast of pregnancy. In this study the effects of synthetic glucocorticoid dexamethasone were inves ti gated in both JAr and JEG-3 cell lines by the MTT test, cell based ELISA, and the cell adhesion and migration tests. Viable cell number/cell proliferation of JAr cells was signi ficantly increased after treatment with 0.1 and 1 nmol/L of dexamethasone, while proliferation of JEG-3 cells was signi ficantly increased after treatment in the whole concentration range of dexamethasone (0.1-100 nmol/L). Galectin-1 in JAr cells was modulated by dexamethasone, which mildly, but significantly decreased production at low concentrations (0.1 and 1 nmol/L). In JEG-3 cells pro duction of galectin-1 was significantly decreased only after treat ment with 100 nmol/L of dexamethasone. Cell adhe sion of JEG-3 was significantly increased in the presence of lacto se, an inhibitory sugar for gal-1, while dexamethasone indu ced decrease of JEG-3 cell migration. These findings have shown that dexamethasone may affect proliferation, gal-1 production and cell migration, in a cell line specific manner. These data suggest that glucocorticoid treatment in vivo might have the potential to affect cell functions in cho riocarcinoma.

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Section: Discussionsupporting

confidence: 49%

Choriocarcinoma cell line Response to Dexamethasone

Bojić‐Trbojević¹,

Kolundžić²,

Petronijević³

et al. 2010

Journal of Medical Biochemistry

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“…Cytotrophoblasts were initially isolated from approximately 45 g of human villous tissue at term following trypsin digestion and centrifugation on Percoll gradients. We have previously employed this procedure [18] which is a modification of those developed by Kliman and colleagues [19] and Douglas and King [20]. Briefly, fresh placental tissue fragments were finely chopped, washed with saline, and treated with trypsin and DNAase.…”

Section: Cell Culturementioning

confidence: 99%

“…Under these conditions we and others have found that we obtain a CT purity of approximately 90-95%, with the major contaminant being FIBs (∼5%) and immune cells (1%) [18,19,21]. Immunopurification was carried out to remove these cell types using procedures as we have previously described [18]. One hundred microliters each of immunomagnetic microspheres conjugated to mouse anti-human CD-45 (immune cell marker) and CD-9 (fibroblast and extravillous trophoblast marker) obtained from Clemente Associates Inc. (Madison, CT) were added to approximately 3 X 10 8 cells.…”

Section: Cell Culturementioning

confidence: 99%

“…FCS medium. Under these conditions we and others have found that we obtain a CT purity of approximately 90-95%, with the major contaminant being FIBs (∼5%) and immune cells (1%) [18,19,21]. Immunopurification was carried out to remove these cell types using procedures as we have previously described [18].…”

Section: Cell Culturementioning

confidence: 99%

“…Overnight incubation with primary antibody at 4º C was followed by a 1 h incubation with rabbit anti-goat (1:2000), goat anti-mouse (1:15,000), or sheep anti-rabbit (1:5,000) horseradish secondary antibody conjugates (Bio-Rad, Hercules, CA). Proteins were detected using an enhanced chemiluminescence reagent (SuperSignal West Femto Maximum Sensitivity Substrate, Pierce, Rockford, IL) as we have previously described [18]. The Western blot in figure 3 depicts results obtained for mFIBs and SCTs isolated from 3 different placentas.…”

Section: Western Blottingmentioning

confidence: 99%

See 2 more Smart Citations

Cell Type-specific Expression and Function of Toll-like Receptors 2 and 4 in Human Placenta: Implications in Fetal Infection

Krikun

Abrahams

et al. 2007

Placenta

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Placental infection is associated with adverse fetal outcomes. Toll-like receptors (TLRs) are critical regulators of the innate immune response based on their ability to recognize and respond to pathogenassociated molecular patterns expressed by microbes. To date, cell-type specific expression and regulation of TLR function in human term placenta remains largely unelucidated. The goal of the current study was to examine the in vivo and in vitro patterns of TLR expression and function in major cell types of term placenta.Immunohistochemical analysis of terminal and stem villi localized TLR-2, which recognizes peptidoglycan (PG) from Gram-positive bacteria, to endothelial cells and macrophages, and to a lesser extent to syncytiotrophoblast (SCTs) and fibroblasts (FIBs). Staining for TLR-4, the receptor for Gram-negative bacterial lipopolysaccharide (LPS), was most prominent in SCTs and endothelial cells. Results from Western blotting, conventional, and quantitative PCR (qRTPCR) analyses using protein and mRNA isolated from cultures of SCTs and myofibroblasts (mFIBs) revealed that SCTs expressed TLR-2 and TLR-4, whereas mFIBs expressed only TLR-4. In addition, qRTPCR showed that LPS treatment increased TLR-2 expression in SCTs, indicating that infection with Gramnegative bacteria may enhance innate immune responses in placenta toward a broad range of microorganisms. In addition, treatment with LPS increased IL-8 levels in both SCTs and mFIBs, whereas PG treatment only stimulated IL-8 levels in SCTs. Our results indicate that there exist cell type-specific patterns of TLR function in placenta which likely regulate innate immune response at the maternal-fetal interface.

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Monoclonal antibody FDC-6 exhibits binding to human plasma fibronectin: A caveat for cervicovaginal oncofetal fibronectin testing?

Feinberg

Wang

1994

American Journal of Obstetrics and Gynecology

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Steroid regulation of oncofetal fibronectin expression in human cytotrophoblasts

Cited by 44 publications

References 26 publications

Choriocarcinoma cell line Response to Dexamethasone

Choriocarcinoma cell line Response to Dexamethasone

Cell Type-specific Expression and Function of Toll-like Receptors 2 and 4 in Human Placenta: Implications in Fetal Infection

Monoclonal antibody FDC-6 exhibits binding to human plasma fibronectin: A caveat for cervicovaginal oncofetal fibronectin testing?

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