scholarly journals Enhancement of Cortisol-Induced 11β-Hydroxysteroid dehydrogenase Type 1 Expression by Interleukin 1β in Cultured Human Chorionic Trophoblast Cells

Endocrinology ◽  
2006 ◽  
Vol 147 (5) ◽  
pp. 2490-2495 ◽  
Author(s):  
Wenjiao Li ◽  
Lu Gao ◽  
Yan Wang ◽  
Tao Duan ◽  
Leslie Myatt ◽  
...  
Keyword(s):  
Gene Expression ◽  
Phospholipase A2 ◽  
Mrna Expression ◽  
Reporter Gene ◽  
Reductase Activity ◽  
Hydroxysteroid Dehydrogenase ◽  
Cytosolic Phospholipase A2 ◽  
Trophoblast Cells ◽  
Cytosolic Phospholipase

Chorion is the most abundant site of 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1) expression within intrauterine tissues. It is important to study the regulation of 11β-HSD1 expression in the chorion in terms of local cortisol production during pregnancy. Using real-time PCR and enzyme activity assay, we found that cortisol (1 μm) and IL-1β (10 ng/ml) for 24 h significantly increased 11β-HSD1 mRNA expression and reductase activity in cultured human chorionic trophoblasts. A further significant increase of 11β-HSD1 mRNA expression and reductase activity was observed with cotreatment of cortisol and IL-1β. To explore the mechanism of induction, 11β-HSD1 promoter was cloned into pGL3 plasmid expressing a luciferase reporter gene. By transfecting the constructed vector into WISH cells, an amnion-derived cell line, we found that cortisol (1 μm) or IL-1β (10 ng/ml) significantly increased reporter gene expression. Likewise, an additional increase in reporter gene expression was observed with cotreatment of cortisol and IL-β. To explore the physiological significance of 11β-HSD1 induction in the chorion, we studied the effect of cortisol on cytosolic phospholipase A2 and cyclooxygenase 2 expression. We found that treatment of chorionic trophoblast cells with cortisol (1 μm) induced both cytosolic phospholipase A2 and cyclooxygenase 2 mRNA expression. We conclude that cortisol up-regulates 11β-HSD1 expression through induction of promoter activity, and the effect was enhanced by IL-1β, suggesting that more biologically active glucocorticoids could be generated in the fetal membranes in the presence of infection, which may consequently feed forward in up-regulation of prostaglandin synthesis.

scholarly journals A novel selective 11β-hydroxysteroid dehydrogenase type 1 inhibitor prevents human adipogenesis

2008 ◽  
Vol 197 (2) ◽  
pp. 297-307 ◽  
Author(s):  
I J Bujalska ◽  
L L Gathercole ◽  
J W Tomlinson ◽  
C Darimont ◽  
J Ermolieff ◽  
...  
Keyword(s):  
Adipose Tissue ◽  
Mrna Expression ◽  
Adipocyte Differentiation ◽  
Reductase Activity ◽  
Primary Cultures ◽  
Hydroxysteroid Dehydrogenase ◽  
Novel Therapy ◽  
Fatty Acid Binding ◽  
Glucose Output

Glucocorticoid excess increases fat mass, preferentially within omental depots; yet circulating cortisol concentrations are normal in most patients with metabolic syndrome (MS). At a pre-receptor level, 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1) activates cortisol from cortisone locally within adipose tissue, and inhibition of 11β-HSD1 in liver and adipose tissue has been proposed as a novel therapy to treat MS by reducing hepatic glucose output and adiposity. Using a transformed human subcutaneous preadipocyte cell line (Chub-S7) and human primary preadipocytes, we have defined the role of glucocorticoids and 11β-HSD1 in regulating adipose tissue differentiation. Human cells were differentiated with 1.0 μM cortisol (F), or cortisone (E) with or without 100 nM of a highly selective 11β-HSD1 inhibitor PF-877423. 11β-HSD1 mRNA expression increased across adipocyte differentiation (P<0.001, n=4), which was paralleled by an increase in 11β-HSD1 oxo-reductase activity (from nil on day 0 to 5.9±1.9 pmol/mg per h on day 16, P<0.01, n=7). Cortisone enhanced adipocyte differentiation; fatty acid-binding protein 4 expression increased 312-fold (P<0.001) and glycerol-3-phosphate dehydrogenase 47-fold (P<0.001) versus controls. This was abolished by co-incubation with PF-877423. In addition, cellular lipid content decreased significantly. These findings were confirmed in the primary cultures of human subcutaneous preadipocytes. The increase in 11β-HSD1 mRNA expression and activity is essential for the induction of human adipogenesis. Blocking adipogenesis with a novel and specific 11β-HSD1 inhibitor may represent a novel approach to treat obesity in patients with MS.

scholarly journals Endothelin-1 stimulates cytosolic phospholipase A2 activity and gene expression in rat glomerular mesangial cells

1994 ◽  
Vol 46 (6) ◽  
pp. 1644-1652 ◽  
Author(s):  
Herbert Schramek ◽  
Yizheng Wang ◽  
Martha Konieczkowski ◽  
Michael S. Simonson ◽  
Michael J. Dunn
Keyword(s):  
Gene Expression ◽  
Phospholipase A2 ◽  
Mesangial Cells ◽  
Cytosolic Phospholipase A2 ◽  
Glomerular Mesangial Cells ◽  
Endothelin 1 ◽  
Cytosolic Phospholipase ◽  
Phospholipase A2 Activity

Angiotensin II Type 1 Receptor Blocker Telmisartan Reduces Cerebral Infarct Volume and Peri-infarct Cytosolic Phospholipase A2 Level in Experimental Stroke

2009 ◽  
Vol 26 (12) ◽  
pp. 2355-2364 ◽  
Author(s):  
Tomonori Kobayashi ◽  
Takakazu Kawamata ◽  
Noriyuki Shibata ◽  
Yoshikazu Okada ◽  
Makio Kobayashi ◽  
...  
Keyword(s):  
Angiotensin Ii ◽  
Phospholipase A2 ◽  
Infarct Volume ◽  
Cytosolic Phospholipase A2 ◽  
Cerebral Infarct ◽  
Receptor Blocker ◽  
Experimental Stroke ◽  
Cytosolic Phospholipase

scholarly journals Cytosolic Phospholipase A2 Alpha Regulates TLR Signaling and Migration in Metastatic 4T1 Cells

2019 ◽  
Vol 20 (19) ◽  
pp. 4800 ◽  
Author(s):  
Hanna Maja Tunset ◽  
Astrid Jullumstrø Feuerherm ◽  
Linn-Karina Myrland Selvik ◽  
Berit Johansen ◽  
Siver Andreas Moestue
Keyword(s):  
Gene Expression ◽  
Phospholipase A2 ◽  
Cell Line ◽  
Enzyme Inhibition ◽  
Cell Lines ◽  
Cytosolic Phospholipase A2 ◽  
Line Pair ◽  
Type I ◽  
Cytosolic Phospholipase ◽  
4T1 Cells

Metastatic disease is the leading cause of death in breast cancer patients. Disrupting the cancer cell’s ability to migrate may be a strategy for hindering metastasis. Cytosolic phospholipase A2 α (cPLA2α), along with downstream proinflammatory and promigratory metabolites, has been implicated in several aspects of tumorigenesis, as well as metastasis, in various types of cancer. In this study, we aim to characterize the response to reduced cPLA2α activity in metastatic versus non-metastatic cells. We employ an isogenic murine cell line pair displaying metastatic (4T1) and non-metastatic (67NR) phenotype to investigate the role of cPLA2α on migration. Furthermore, we elucidate the effect of reduced cPLA2α activity on global gene expression in the metastatic cell line. Enzyme inhibition is achieved by using a competitive pharmacological inhibitor, cPLA2α inhibitor X (CIX). Our data show that 4T1 expresses significantly higher cPLA2α levels as compared to 67NR, and the two cell lines show different sensitivity to the CIX treatment with regards to metabolism and proliferation. Inhibition of cPLA2α at nontoxic concentrations attenuates migration of highly metastatic 4T1 cells, but not non-metastatic 67NR cells. Gene expression analysis indicates that processes such as interferon type I (IFN-I) signaling and cell cycle regulation are key processes regulated by cPLA2a in metastatic 4T1 cells, supporting the findings from the biological assays. This study demonstrates that two isogenic cancer cell lines with different metastatic potential respond differently to reduced cPLA2α activity. In conclusion, we argue that cPLA2α is a potential therapeutic target in cancer and that enzyme inhibition may inhibit metastasis through an anti-migratory mechanism, possibly involving Toll-like receptor signaling and type I interferons.

scholarly journals Regulation of 11beta-hydroxysteroid dehydrogenase type 1 gene expression by LH and interleukin-1beta in cultured rat granulosa cells

1999 ◽  
Vol 163 (3) ◽  
pp. 417-423 ◽  
Author(s):  
M Tetsuka ◽  
LC Haines ◽  
M Milne ◽  
GE Simpson ◽  
SG Hillier
Keyword(s):  
Gene Expression ◽  
Mrna Expression ◽  
Granulosa Cells ◽  
Hydroxysteroid Dehydrogenase ◽  
Releasing Hormone ◽  
Cell Monolayers ◽  
Interleukin 1Beta ◽  
Lh Releasing Hormone

Granulosa cells from preovulatory follicles show increased expression of 11beta-hydroxysteroid dehydrogenase type 1 (11betaHSD1) at the time of ovulation. As ovulation may be an inflammatory process, this may be a mechanism of local enhancement of the activity of anti-inflammatory glucocorticoids. In this study, we examined direct effects of LH, the proinflammatory cytokine, interleukin-1beta (IL-1beta), and pharmacological activators of protein kinase A (PKA) (forskolin and dibutyryl (db) cAMP) and PKC (LH-releasing hormone and phorbol 12-myristate 13-acetate (PMA)) signalling on the expression of 11betaHSD1 mRNA in vitro. Granulosa cells from immature female rat ovaries were cultured (pretreatment) in serum-free medium 199 containing recombinant human (rh) FSH (1 ng/ml) for 48 h to induce responsiveness to LH. Cell monolayers were then washed and cultured (test treatment) for a further 12 h in the presence of rhLH (0-100 ng/ml), IL-1beta (0-50 ng/ml), or both. Total RNA was extracted from granulosa cell monolayers and taken for quantitative ribonuclease protection analysis of 11betaHSD1 mRNA. The low level of 11betaHSD1 mRNA detectable in unstimulated (control) cultures was increased approximately twofold by the 48-h pretreatment with rhFSH. Subsequent exposure to rhLH (1-100 ng/ml) for a further 12 h dose-dependently increased 11betaHSD1 mRNA expression by an additional two- to threefold. Forskolin (10 microM), db-cAMP (2 mM), LH-releasing hormone (LHRH; 1 microM) and PMA (200 nM) were also stimulatory. IL-1beta (0.05-50 ng/ml) stimulated 11betaHSD1 mRNA expression in a dose-related manner, both in the absence and in the presence of rhLH (3 ng/ml). The interaction between IL-1beta (5 ng/ml) and rhLH (3 ng/ml) was additive. Co-treatment with a 50-fold excess of IL-1 receptor antagonist fully reversed the action of IL-1beta. We conclude that 11betaHSD1 mRNA expression in functionally mature granulosa cells is directly stimulated by gonadotrophins and IL-1beta in vitro, potentially involving post-receptor signalling via PKA- and PKC-mediated pathways. Thus both LH and IL-1beta may serve physiological roles in the upregulation of 11betaHSD1 gene expression by granulosa cells in ovulatory follicles.

scholarly journals Cytosolic phospholipase A2 gene expression in rat mesangial cells is regulated post-transcriptionally

1994 ◽  
Vol 304 (2) ◽  
pp. 417-422 ◽  
Author(s):  
A Tay ◽  
P Maxwell ◽  
Z G Li ◽  
H Goldberg ◽  
K Skorecki
Keyword(s):  
Gene Expression ◽  
Growth Factor ◽  
Phospholipase A2 ◽  
Mesangial Cells ◽  
Luciferase Activity ◽  
Cytosolic Phospholipase A2 ◽  
Mrna Levels ◽  
Glomerular Mesangial Cells ◽  
Coding Region ◽  
Cytosolic Phospholipase

Cytosolic phospholipase A2 (cPLA2) is thought to be the rate-limiting enzyme in the arachidonic acid/eicosanoid cascade. The ability of various agonists to increase steady-state cPLA2 mRNA levels has previously been reported. The current study delineates the contributions of transcriptional and post-transcriptional processes to the regulation of cPLA2 gene expression in response to a variety of agonists in cultured rat glomerular mesangial cells. Epidermal growth factor, platelet-derived growth factor, serum and phorbol myristate acetate all increase the half-life of cPLA2 mRNA transcripts, indicating a role for post-transcriptional modulation of gene expression. The presence of three ATTTA motifs in the 3′ untranslated region (3′UTR) of the rat cPLA2 cDNA is ascertained. Heterologous expression of chimeric constructs with different 3′UTRs ligated into the 3′ end of the luciferase coding region reveals that the presence of the cPLA2 3′UTR results in reduced luciferase activity compared with constructs without the cPLA2 3′UTR. Furthermore, the luciferase activity in the constructs with the cPLA2 3′UTR is increased in response to the same agonists which stabilize endogenous cPLA2 mRNA. A negligible effect of these agonists on transcriptional control of cPLA2 is evident using promoter-reporter constructs expressed in transient and stable transfectants. Taken together, these results indicate predominant post-transcriptional regulation of cPLA2 mRNA levels.

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