title: Inhibitory effects of prostaglandin E2 on collagen synthesis and cell proliferation in human stellate cells from pancreatic head adenocarcinoma.
abstract: BACKGROUND
  Several studies have described an increased cyclooxygenase-2 (COX-2) expression in pancreatic cancer, but the role of COX-2 in tumour development and progression is not clear. The aim of the present study was to examine expression of COX-2 in cancer cells and stromal cells in pancreatic cancer specimens, and to explore the role of PGE2 in pancreatic stellate cell proliferation and collagen synthesis.
  
  
  METHODS
  Immunohistochemistry and immunofluorescence was performed on slides from whole sections of tissue blocks using antibodies against COX-2 and α-smooth muscle actin (αSMA). Pancreatic stellate cells (PSC) were isolated from surgically resected tumour tissue by the outgrowth method. Cells were used between passages 4 and 8. Collagen synthesis was determined by [(3)H]-proline incorporation, or by enzyme immunoassay measurement of collagen C-peptide. DNA synthesis was measured by incorporation of [(3)H]-thymidine in DNA. Cyclic AMP (cAMP) was determined by radioimmunoassay. Collagen 1A1 mRNA was determined by RT-qPCR.
  
  
  RESULTS
  Immunohistochemistry staining showed COX-2 in pancreatic carcinoma cells, but not in stromal cells. All tumours showed positive staining for αSMA in the fibrotic stroma. Cultured PSC expressed COX-2, which could be further induced by interleukin-1β (IL-1β), epidermal growth factor (EGF), thrombin, and PGE2, but not by transforming growth factor-β1 (TGFβ). Indirect coculture with the adenocarcinoma cell line BxPC-3, but not HPAFII or Panc-1, induced COX-2 expression in PSC. Treatment of PSC with PGE2 strongly stimulated cAMP accumulation, mediated by EP2 receptors, and also stimulated phosphorylation of extracellular signal-regulated kinase (ERK). Treatment of PSC with PGE2 or forskolin suppressed both TGFβ-stimulated collagen synthesis and PDGF-stimulated DNA synthesis.
  
  
  CONCLUSIONS
  The present results show that COX-2 is mainly produced in carcinoma cells and suggest that the cancer cells are the main source of PGE2 in pancreatic tumours. PGE2 exerts a suppressive effect on proliferation and fibrogenesis in pancreatic stellate cells. These effects of PGE2 are mediated by the cAMP pathway and suggest a role of EP2 receptors.
authors.affiliation: Institute of Clinical Medicine, Faculty of Medicine, University of Oslo, Oslo, Norway. ewa.pomianowska@medisin.uio.no.
authors.forename: Ewa
authors.initials: E
authors.lastname: Pomianowska
authors.forename: Dagny
authors.initials: D
authors.lastname: Sandnes
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Grzyb
authors.forename: Aasa R
authors.initials: AR
authors.lastname: Schjølberg
authors.forename: Monica
authors.initials: M
authors.lastname: Aasrum
authors.forename: Ingun H
authors.initials: IH
authors.lastname: Tveteraas
authors.forename: Vegard
authors.initials: V
authors.lastname: Tjomsland
authors.forename: Thoralf
authors.initials: T
authors.lastname: Christoffersen
authors.forename: Ivar P
authors.initials: IP
authors.lastname: Gladhaug
mesh_terms: D045744:Cell Line, Tumor; D002478:Cells, Cultured; D018920:Coculture Techniques; D003094:Collagen / Q000096:biosynthesis*; D051546:Cyclooxygenase 2 / Q000235:genetics / Q000378:metabolism; D004261:DNA Replication / Q000187:drug effects; D015232:Dinoprostone / Q000494:pharmacology*; D015870:Gene Expression; D006801:Humans; D007150:Immunohistochemistry; D010190:Pancreatic Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology*; D058954:Pancreatic Stellate Cells / Q000187:drug effects* / Q000378:metabolism*
chemical_list: D003094:Collagen; D051546:Cyclooxygenase 2; D015232:Dinoprostone