title: Trefoil factors: tumor progression markers and mitogens via EGFR/MAPK activation in cholangiocarcinoma.
abstract: OBJECTIVE
  To investigate trefoil factor (TFF) gene copy number, mRNA and protein expression as potential biomarkers in cholangiocarcinoma (CCA).
  
  
  METHODS
  TFF mRNA levels, gene copy number and protein expression were determined respectively by quantitative reverse transcription polymerase chain reaction (PCR), quantitative PCR and immunohistochemistry in bile duct epithelium biopsies collected from individuals with CCA, precancerous bile duct dysplasia and from disease-free controls. The functional impact of recombinant human (rh)TFF2 peptide treatment on proliferation and epidermal growth factor receptor (EGFR)/mitogen-activated protein kinase (MAPK) signaling was assessed in the CCA cell line, KMBC, by viable cell counting and immunoblotting, respectively.
  
  
  RESULTS
  TFF1, TFF2 and TFF3 mRNA expression was significantly increased in CCA tissue compared to disease-free controls, and was unrelated to gene copy number. TFF1 immunoreactivity was strongly increased in both dysplasia and CCA, whereas TFF2 immunoreactivity was increased only in CCA compared to disease-free controls. By contrast, TFF3 immunoreactivity was moderately decreased in dysplasia and further decreased in CCA. Kaplan-Meier analysis found no association of TFF mRNA, protein and copy number with age, gender, histological subtype, and patient survival time. Treatment of KMBC cells with rhTFF2 stimulated proliferation, triggered phosphorylation of EGFR and downstream extracellular signal related kinase (ERK), whereas co-incubation with the EGFR tyrosine kinase inhibitor, PD153035, blocked rhTFF2-dependent proliferation and EGFR/ERK responses.
  
  
  CONCLUSIONS
  TFF mRNA/protein expression is indicative of CCA tumor progression, but not predictive for histological sub-type or survival time. TFF2 is mitogenic in CCA via EGFR/MAPK activation.
authors.affiliation: Graduate School, Centre for Research and Development of Medical Diagnostic Laboratories, Faculty of Associated Medical Sciences, Khon Kaen University, Khon Kaen 40002, Thailand.
authors.forename: Kanuengnuch
authors.initials: K
authors.lastname: Kosriwong
authors.forename: Trevelyan R
authors.initials: TR
authors.lastname: Menheniott
authors.forename: Andrew S
authors.initials: AS
authors.lastname: Giraud
authors.forename: Patcharee
authors.initials: P
authors.lastname: Jearanaikoon
authors.forename: Banchob
authors.initials: B
authors.lastname: Sripa
authors.forename: Temduang
authors.initials: T
authors.lastname: Limpaiboon
mesh_terms: D001650:Bile Duct Neoplasms / Q000201:enzymology* / Q000235:genetics / Q000401:mortality / Q000473:pathology; D001653:Bile Ducts, Intrahepatic / Q000201:enzymology* / Q000473:pathology; D001706:Biopsy; D016022:Case-Control Studies; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D016009:Chi-Square Distribution; D018281:Cholangiocarcinoma / Q000201:enzymology* / Q000235:genetics / Q000401:mortality / Q000473:pathology; D018450:Disease Progression; D004789:Enzyme Activation; D066246:ErbB Receptors / Q000378:metabolism*; D018628:Gene Dosage; D006801:Humans; D007150:Immunohistochemistry; D053208:Kaplan-Meier Estimate; D016014:Linear Models; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism*; D010455:Peptides / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D011379:Prognosis; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D000071163:Trefoil Factor-1; D000071164:Trefoil Factor-2; D000071165:Trefoil Factor-3; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D010455:Peptides; D012333:RNA, Messenger; D011994:Recombinant Proteins; C051838:TFF1 protein, human; C000604007:TFF2 protein, human; C082495:TFF3 protein, human; D000071163:Trefoil Factor-1; D000071164:Trefoil Factor-2; D000071165:Trefoil Factor-3; D025521:Tumor Suppressor Proteins; C512478:EGFR protein, human; D066246:ErbB Receptors; D020928:Mitogen-Activated Protein Kinases