title: Cellular senescence involves an intracrine prostaglandin E2 pathway in human fibroblasts.
abstract: Cyclooxygenase 2 and release of prostaglandin E2 are involved in many responses including inflammation and are upregulated during cellular senescence. However, little is known about the role of lipid inflammatory mediators in senescence. Here, we investigated the mechanism by which the COX-2/PGE2 axis induces senescence. Using the NS398 specific inhibitor of COX-2, we provide evidence that reactive oxygen species by-produced by the COX-2 enzymatic activity are negligible in front of the total senescence-associated oxidative stress. We therefore investigated the role of PGE2 by invalidating the PGE2 synthases downstream of COX-2, or the specific PGE2 receptors, or by applying PGE2 or specific agonists or antagonists. We evaluated the effect on senescence by evaluating the senescence-associated proliferation arrest, the percentage of senescence-associated beta-galactosidase-positive cells, and the expression of senescent molecular markers such as IL-6 and MCP1. We show that PGE2 acting on its EP specific receptors is able to induce both the onset of senescence and the maintenance of the phenotype. It did so only when the PGE2/lactate transporter activity was enhanced, indicating that PGE2 acts on senescence more via the pool of intracellular EP receptors than via those localized at the cell surface. Treatment with agonists, antagonists and silencing of the EP receptors by siRNA revealed that EP3 was the most involved in transducing the intracrine effects of PGE2. Immunofluorescence experiments confirmed that EP3 was more localized in the cytoplasm than at the cell surface. Taken together, these results suggest that COX-2 contributes to the establishment and maintenance of senescence of normal human fibroblasts via an independent-ROS and a dependent-PGE2/EPs intracrine pathway.
authors.affiliation: CNRS, UMR8161, Institut de Biologie de Lille, 1 rue Calmette, F-59021 Lille Cedex, France.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Martien
authors.forename: Olivier
authors.initials: O
authors.lastname: Pluquet
authors.forename: Chantal
authors.initials: C
authors.lastname: Vercamer
authors.forename: Nicolas
authors.initials: N
authors.lastname: Malaquin
authors.forename: Nathalie
authors.initials: N
authors.lastname: Martin
authors.forename: Karo
authors.initials: K
authors.lastname: Gosselin
authors.forename: Albin
authors.initials: A
authors.lastname: Pourtier
authors.forename: Corinne
authors.initials: C
authors.lastname: Abbadie
mesh_terms: D002460:Cell Line; D016922:Cellular Senescence; D051546:Cyclooxygenase 2 / Q000378:metabolism*; D020405:Dermis / Q000166:cytology; D015232:Dinoprostone / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D018384:Oxidative Stress; D017382:Reactive Oxygen Species / Q000378:metabolism; D058307:Receptors, Prostaglandin E, EP2 Subtype / Q000378:metabolism; D058308:Receptors, Prostaglandin E, EP3 Subtype / Q000378:metabolism; D015398:Signal Transduction
chemical_list: D017382:Reactive Oxygen Species; D058307:Receptors, Prostaglandin E, EP2 Subtype; D058308:Receptors, Prostaglandin E, EP3 Subtype; D051546:Cyclooxygenase 2; D015232:Dinoprostone