title: The induction of S100p expression by the Prostaglandin E₂ (PGE₂)/EP4 receptor signaling pathway in colon cancer cells.
abstract: BACKGROUND
  Prostaglandin E₂ (PGE₂) levels are frequently elevated in colorectal carcinomas. PGE₂ is perceived via four transmembrane G protein coupled receptors (EP1-4), among which the EP4 receptor is most relevant. PGE₂/EP4-receptor interaction activates CREB via the ERK/MEK pathway. However, the downstream target genes activated by this pathway remained to be investigated.
  
  
  RESULTS
  Here, we have identified S100P (an EF-hand calcium binding protein) as a novel downstream target. We show by realtime RT-PCR that S100P mRNA levels are elevated in 14/17 (82%) colon tumor tissues as compared to paired adjacent normal colonic tissues. S100P expression is stimulated in the presence of PGE₂ in a time dependent manner at mRNA and protein levels in colon, breast and pancreatic cancer cells. Pharmacological and RNAi-mediated inhibition of the EP4 receptor attenuates PGE₂-dependent S100P mRNA induction. RNA(i)-mediated knockdown of CREB inhibits endogenous S100P expression. Furthermore, using luciferase reporter analysis and EMSA we show that mutation and/or deletion of the CRE sequence within the S100P promoter abolished PGE₂-mediated transcriptional induction. Finally, we demonstrate that RNA(i)-mediated knockdown of S100P compromised invadopodia formation, colony growth and motility of colon cancer cells. Interestingly, endogenous knock down of S100P decreases ERK expression levels, suggesting a role for ERK in regulating S100P mediated cell growth and motility.
  
  
  CONCLUSIONS
  Together, our findings show for the first time that S100P expression is regulated by PGE₂/EP4-receptor signaling and may participate in a feedback signaling that perpetuates tumor cell growth and migration. Therefore, our data suggest that dysregulated S100P expression resulting from aberrant PGE₂/EP4 receptor signaling may have important consequences relevant to colon cancer pathogenesis.
authors.affiliation: Department of Pathology, Arizona Cancer Center, Tucson, USA.
authors.forename: Anupama
authors.initials: A
authors.lastname: Chandramouli
authors.forename: Melania E
authors.initials: ME
authors.lastname: Mercado-Pimentel
authors.forename: Anthony
authors.initials: A
authors.lastname: Hutchinson
authors.forename: Adriana
authors.initials: A
authors.lastname: Gibadulinová
authors.forename: Erik R
authors.initials: ER
authors.lastname: Olson
authors.forename: Sally
authors.initials: S
authors.lastname: Dickinson
authors.forename: Reneé
authors.initials: R
authors.lastname: Shañas
authors.forename: Jennifer
authors.initials: J
authors.lastname: Davenport
authors.forename: Janae
authors.initials: J
authors.lastname: Owens
authors.forename: Achyut K
authors.initials: AK
authors.lastname: Bhattacharyya
authors.forename: John W
authors.initials: JW
authors.lastname: Regan
authors.forename: Silvia
authors.initials: S
authors.lastname: Pastorekova
authors.forename: Thiruvengadam
authors.initials: T
authors.lastname: Arumugam
authors.forename: Craig D
authors.initials: CD
authors.lastname: Logsdon
authors.forename: Mark A
authors.initials: MA
authors.lastname: Nelson
mesh_terms: D015153:Blotting, Western; D002135:Calcium-Binding Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D003110:Colonic Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D003114:Colony-Forming Units Assay; D017362:Cyclic AMP Response Element-Binding Protein / Q000235:genetics / Q000378:metabolism; D015232:Dinoprostone / Q000235:genetics / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay; D048049:Extracellular Signal-Regulated MAP Kinases / Q000235:genetics / Q000378:metabolism; D006801:Humans; D020929:Mitogen-Activated Protein Kinase Kinases / Q000235:genetics / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D009154:Mutation / Q000235:genetics; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D058309:Receptors, Prostaglandin E, EP4 Subtype / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D014407:Tumor Cells, Cultured
chemical_list: C115163:CREB1 protein, human; D002135:Calcium-Binding Proteins; D017362:Cyclic AMP Response Element-Binding Protein; D009363:Neoplasm Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D058309:Receptors, Prostaglandin E, EP4 Subtype; C103003:S100P protein, human; D048049:Extracellular Signal-Regulated MAP Kinases; D020929:Mitogen-Activated Protein Kinase Kinases; D015232:Dinoprostone