Journal Article

PGF2α - FP Receptor Regulation of Interleukin-11 Expression in Endometrial Adenocarcinoma Cells Via the Calcium-Calcineurin Pathway

Anderson, Richard A., Grant, Vivien, Maldonado-Perez, David, Jabbour, Henry N., Sales, Kurt J., Williams, Alistair R.W.

2008

Overview

The pleiotropic cytokine interleukin-11 (IL-11) up-regulates the proliferative and invasive capacity of many human cancer cells in vitro. Co-expression of gp130 and IL-11 receptor alpha (IL-11rα) is necessary for high affinity binding of IL-11 to IL-11rα. Recently we have established a role for prostaglandin F2α (PGF) receptor (FP) signalling in the regulation of tumorigenic genes in endometrial adenocarcinomas. This study investigated the expression of IL-11 and role of PGF-FP receptor signaling in the modulation of IL-11 expression in endometrial adenocarcinoma cells. Human adenocarcinoma biopsies were obtained in accordance with Lothian Research Ethics Committee guidance with informed patient consent. Localisation of IL-11, IL-11rα and gp130 expression was performed by immunohistochemistry. IL-11 mRNA expression was determined by real-time RT-PCR analysis using Ishikawa endometrial adenocarcinoma cells expressing the FP receptor to the levels observed in endometrial adenocarcinomas (FPS cells). IL-11 mRNA expression was significantly elevated in endometrial adenocarcinoma samples compared to normal endometrium (0.4 ± 0.1 vs 0.03 ± 0.01 arbitrary units; p<0.05) and localized with IL-11rα and gp130 in the glandular epithelium of endometrial adenocarcinomas. To investigate PGF-FP receptor signaling to IL-11 in endometrial adenocarcinoma cells, FPS cells were treated with vehicle, 100nM PGF or 100nM PGF in the absence/presence of the FP receptor antagonist AL8810 or chemical inhibitors of protein kinase A (PKA; 4C3MQ), protein kinase C (PKC; RO318220), calcineurin (cyclosporine A), Calcium (EGTA) or NFAT (INCa6). Treatment of FPS cells with PGF for 24 hours significantly elevated the expression of IL-11 mRNA (95.1 ± 6.4 fold above vehicle treated cells, P<0.05). Co-incubation of FPS cells with PGF and the FP receptor antagonist AL8810 (8.4 ± 6.4 fold above vehicle treated cells; P<0.01) or chemical inhibitors of PKC (9.6 ± 4.1 fold; P<0.01), calcineurin (17.7 ± 3.7 fold; P<0.01), calcium (15.8 ± 9.5 fold; P<0.05) or NFAT (18.3 ± 10.2 fold; P<0.01) but not PKA (96.5 ± 32.7 fold; P<0.01) significantly reduced the PGF-mediated increase in IL-11 mRNA expression. These data demonstrate that PGF regulation of IL-11 mRNA is mediated via the activation of PKC and the calcium-calcineurin pathway and suggest a possible mechanism for control of endometrial tumorigenesis by PGF-FP receptor via activation of signaling to cytokines such as IL-11.

• Publisher: Oxford University Press
• Subject: Adenocarcinoma / Biopsy / Calcineurin / Calcium / Cancer / Cyclosporins / Cytokines / Endometrium / Epithelium / Gene expression / Gene regulation / Glycoprotein gp130 / Immunohistochemistry / Interleukin 11 / Kinases / NF-AT protein / Prostaglandin F2a / Prostaglandins / Protein kinase A / Protein kinase C / Signal transduction / Tumorigenesis