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    Proneoplastic effects of PGE2 mediated by EP4 receptor in colorectal cancer.

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    Authors
    Doherty, Glen A
    Byrne, Sinead M
    Molloy, Eamonn S
    Malhotra, Vikrum
    Austin, Sandra C
    Kay, Elaine W
    Murray, Frank E
    Fitzgerald, Desmond J
    Affiliation
    Molecular Medicine Group, Conway Institute of Biomolecular and Biomedical Research, University College Dublin, Dublin, Ireland. glen_doherty@hotmail.com
    Issue Date
    2009
    MeSH
    Cell Line, Tumor
    Cell Membrane
    Cell Proliferation
    Colorectal Neoplasms
    Cyclin-Dependent Kinase Inhibitor p21
    Cyclooxygenase 2
    Dinoprostone
    Flow Cytometry
    Gene Expression Regulation, Neoplastic
    Humans
    Immunohistochemistry
    Precancerous Conditions
    Receptor, Epidermal Growth Factor
    Receptors, Prostaglandin E
    Reverse Transcriptase Polymerase Chain Reaction
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    Citation
    Proneoplastic effects of PGE2 mediated by EP4 receptor in colorectal cancer. 2009, 9:207 BMC Cancer
    Journal
    BMC cancer
    URI
    http://hdl.handle.net/10147/94218
    DOI
    10.1186/1471-2407-9-207
    PubMed ID
    19558693
    Abstract
    BACKGROUND: Prostaglandin E2 (PGE2) is the major product of Cyclooxygenase-2 (COX-2) in colorectal cancer (CRC). We aimed to assess PGE2 cell surface receptors (EP 1-4) to examine the mechanisms by which PGE2 regulates tumour progression. METHODS: Gene expression studies were performed by quantitative RT-PCR. Cell cycle was analysed by flow cytometry with cell proliferation quantified by BrdU incorporation measured by enzyme immunoassay. Immunohistochemistry was employed for expression studies on formalin fixed paraffin embedded tumour tissue. RESULTS: EP4 was the most abundant subtype of PGE2 receptor in HT-29 and HCA7 cells (which show COX-2 dependent PGE2 generation) and was consistently the most abundant transcript in human colorectal tumours (n = 8) by qRT-PCR (ANOVA, p = 0.01). G0/G1 cell cycle arrest was observed in HT-29 cells treated with SC-236 5 microM (selective COX-2 inhibitor) for 24 hours (p = 0.02), an effect abrogated by co-incubation with PGE2 (1 microM). G0/G1 arrest was also seen with a specific EP4 receptor antagonist (EP4A, L-161982) (p = 0.01). Treatment of HT-29 cells with either SC-236 or EP4A caused reduction in intracellular cAMP (ANOVA, p = 0.01). Early induction in p21WAF1/CIP1 expression (by qRT-PCR) was seen with EP4A treatment (mean fold increase 4.4, p = 0.04) while other genes remained unchanged. Similar induction in p21WAF1/CIP1 was also seen with PD153025 (1 microM), an EGFR tyrosine kinase inhibitor, suggesting EGFR transactivation by EP4 as a potential mechanism. Additive inhibition of HCA7 proliferation was observed with the combination of SC-236 and neutralising antibody to amphiregulin (AR), a soluble EGFR ligand. Concordance in COX-2 and AR localisation in human colorectal tumours was noted. CONCLUSION: COX-2 regulates cell cycle transition via EP4 receptor and altered p21WAF1/CIP1 expression. EGFR pathways appear important. Specific targeting of the EP4 receptor or downstream targets may offer a safer alternative to COX-2 inhibition in the chemoprevention of CRC.
    Language
    en
    ISSN
    1471-2407
    ae974a485f413a2113503eed53cd6c53
    10.1186/1471-2407-9-207
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