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Prostaglandin E2-EP4 receptor signalling promotes tumorigenic behaviour of HT-29 human colorectal cancer cells

Oncogene volume 26, pages 3006–3019 (2007)Cite this article

Abstract

The predominant product of cyclooxygenase (COX) activity in the colon, prostaglandin (PG) E2 promotes intestinal tumorigenesis. Expression of the PGE2 receptor EP4 is upregulated during colorectal carcinogenesis. Therefore, we investigated the role of elevated PGE2-EP4 receptor signalling in the protumorigenic activity of PGE2 by increasing EP4 receptor expression in HT-29 human colorectal cancer (CRC) cells (HT-29-EP4) by stable transfection. Elevated PGE2-induced EP4 receptor activity in HT-29 cells increased resistance to spontaneous apoptosis and promoted anchorage-independent growth, but had no effect on proliferation of HT-29-EP4 cells. EP4 receptor activation by PGE2 in HT-29-EP4 cells also led to development of fluid-filled cysts, which was associated with increased tight junction protein (occludin and zonula occludens-1) expression. Overexpression of the EP4 receptor in HT-29 cells led to basal EP4 receptor signalling in the absence of exogenous PGE2, which was explained by autocrine activity of endogenous, COX-2-derived PGE2 and constitutive, ligand-independent EP4 receptor activity. The predominant signalling pathway mediating antiapoptotic activity downstream of PGE2-EP4 receptor activation in HT-29-EP4 cells was elevation of cyclic adenosine monophosphate (cAMP) levels, which was associated with phosphorylation of cAMP-response element binding protein. EP4 receptor activation led to a small increase in phosphorylated extracellular signal-regulated kinase (ERK) 2 protein levels but inhibition of ERK phosphorylation did not abrogate the antiapoptotic activity of PGE2. However, PGE2-EP4 receptor signalling did not lead to trans-activation of the epidermal growth factor receptor in HT-29 cells. Inhibition of protumorigenic PGE2-EP4 receptor signalling represents a potential strategy for anti-CRC therapy that may avoid the toxicity associated with systemic COX inhibition.

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Acknowledgements

We thank Dr John Regan for the kind gift of the pCEP4-EP4 plasmid. Dr Ewan Morrison gave valuable help with time-lapse microscopy. Dr Valerie Speirs provided human EP4 receptor primers for real-time PCR. This work was funded by the Association for International Cancer Research. MA Hull is a Medical Research Council (UK) Senior Clinical Fellow, work in his laboratory is also funded by the World Cancer Research Fund and Yorkshire Cancer Research.

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  1. Section of Molecular Gastroenterology, Leeds Institute of Molecular Medicine, University of Leeds, St James's University Hospital, Leeds, LS9 7TF, UK

    G Hawcroft, C W S Ko & M A Hull

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  1. G Hawcroft
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Correspondence to G Hawcroft.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).

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Hawcroft, G., Ko, C. & Hull, M. Prostaglandin E2-EP4 receptor signalling promotes tumorigenic behaviour of HT-29 human colorectal cancer cells. Oncogene 26, 3006–3019 (2007). https://doi.org/10.1038/sj.onc.1210113

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