title: Distinct antifibrogenic effects of erlotinib, sunitinib and sorafenib on rat pancreatic stellate cells.
abstract: OBJECTIVE
  To study if three clinically available small molecule kinase inhibitors (SMI), erlotinib, sunitinib and sorafenib, exert antifibrogenic effects on pancreatic stellate cells (PSC) and analyze the basis of their action.
  
  
  METHODS
  Cultured rat PSC were exposed to SMI. Cell proliferation and viability were assessed employing 5-bromo-2'-deoxyuridine incorporation assay and flow cytometry, respectively. 2-Deoxy-2-[(18)F] fluoroglucose ((18)F-FDG) uptake was measured to study metabolic activity. Exhibition of the myofibroblastic PSC phenotype was monitored by immunofluorescence analysis of α-smooth muscle actin (α-SMA) expression. Levels of mRNA were determined by real-time PCR, while protein expression and phosphorylation were analyzed by immunoblotting. Transforming growth factor-β1 (TGF-β1) levels in culture supernatants were quantified by ELISA.
  
  
  RESULTS
  All three SMI inhibited cell proliferation and (18)F-FDG uptake in a dose-dependent manner and without significant cytotoxic effects. Furthermore, additive effects of the drugs were observed. Immunoblot analysis showed that sorafenib and sunitib, but not erlotinib, efficiently blocked activation of the AKT pathway, while all three drugs displayed little effect on phosphorylation of ERK1/2. Cells treated with sorafenib or sunitinib expressed less interleukin-6 mRNA as well as less collagen type 1 mRNA and protein. Sorafenib was the only drug that also upregulated the expression of matrix metalloproteinase-2 and reduced the secretion of TGF-β1 protein. All three drugs showed insignificant or discordant effects on the mRNA and protein levels of α-SMA.
  
  
  CONCLUSIONS
  The tested SMI, especially sorafenib, exert inhibitory effects on activated PSC, which should be further evaluated in preclinical studies.
authors: Anne Elsner, Falko Lange, Brit Fitzner, Robert Jaster, Department of Medicine II, Division of Gastroenterology, University Medicine Rostock, 18057 Rostock, Germany.
authors: Anne
authors: A
authors: Elsner
authors: Anne Elsner, Falko Lange, Brit Fitzner, Robert Jaster, Department of Medicine II, Division of Gastroenterology, University Medicine Rostock, 18057 Rostock, Germany.
authors: Falko
authors: F
authors: Lange
authors: Anne Elsner, Falko Lange, Brit Fitzner, Robert Jaster, Department of Medicine II, Division of Gastroenterology, University Medicine Rostock, 18057 Rostock, Germany.
authors: Brit
authors: B
authors: Fitzner
authors: Anne Elsner, Falko Lange, Brit Fitzner, Robert Jaster, Department of Medicine II, Division of Gastroenterology, University Medicine Rostock, 18057 Rostock, Germany.
authors: Martin
authors: M
authors: Heuschkel
authors: Anne Elsner, Falko Lange, Brit Fitzner, Robert Jaster, Department of Medicine II, Division of Gastroenterology, University Medicine Rostock, 18057 Rostock, Germany.
authors: Bernd Joachim
authors: BJ
authors: Krause
authors: Anne Elsner, Falko Lange, Brit Fitzner, Robert Jaster, Department of Medicine II, Division of Gastroenterology, University Medicine Rostock, 18057 Rostock, Germany.
authors: Robert
authors: R
authors: Jaster
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism; D000090602:Collagen Type I, alpha 1 Chain; D004305:Dose-Response Relationship, Drug; D000069347:Erlotinib Hydrochloride; D048049:Extracellular Signal-Regulated MAP Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D005786:Gene Expression Regulation; D007211:Indoles / Q000494:pharmacology*; D008297:Male; D020778:Matrix Metalloproteinase 2 / Q000235:genetics / Q000378:metabolism; D009536:Niacinamide / Q000031:analogs & derivatives* / Q000494:pharmacology; D010179:Pancreas / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D058954:Pancreatic Stellate Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D010641:Phenotype; D010671:Phenylurea Compounds / Q000494:pharmacology*; D010766:Phosphorylation; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D051057:Proto-Oncogene Proteins c-akt / Q000037:antagonists & inhibitors / Q000378:metabolism; D011758:Pyrroles / Q000494:pharmacology*; D011799:Quinazolines / Q000494:pharmacology*; D012333:RNA, Messenger / Q000378:metabolism; D011917:Rats, Inbred Lew; D015398:Signal Transduction / Q000187:drug effects; D000077157:Sorafenib; D000077210:Sunitinib; D013997:Time Factors; D053773:Transforming Growth Factor beta1 / Q000235:genetics / Q000378:metabolism
chemical_list: D000199:Actins; D024042:Collagen Type I; D000090602:Collagen Type I, alpha 1 Chain; D000069347:Erlotinib Hydrochloride; D048049:Extracellular Signal-Regulated MAP Kinases; D007211:Indoles; D020778:Matrix Metalloproteinase 2; D009536:Niacinamide; D010671:Phenylurea Compounds; D047428:Protein Kinase Inhibitors; D051057:Proto-Oncogene Proteins c-akt; D011758:Pyrroles; D011799:Quinazolines; D012333:RNA, Messenger; D000077157:Sorafenib; D000077210:Sunitinib; D053773:Transforming Growth Factor beta1; C508962:Tgfb1 protein, rat; C498862:Acta2 protein, rat; C522363:Mmp2 protein, rat
keywords: Erlotinib; Fibrosis; Pancreatic stellate cell; Sorafenib; Sunitinib