title: Tumor necrosis factor-α modulates epithelial mesenchymal transition mediators ZEB2 and S100A4 to promote cholangiocarcinoma progression.
abstract: BACKGROUND
  The epithelial-mesenchymal transition (EMT) process strongly contributes to cancer metastasis. This study was to investigate the alteration of EMT-related proteins (ZEB1, ZEB2 and S100A4) in cholangiocarcinoma (CCA) tissues. The effect of tumor necrosis factor-α (TNF-α) on the expression of those molecules in CCA cells was investigated.
  
  
  METHODS
  The quantitative reverse transcription-polymerase chain reaction (qRT-PCR) assay was used to quantify ZEB1, ZEB2 and S100A4 mRNA levels in 50 CCA tissues and related its expression to clinicopathological data. ZEB2 protein immunostaining was investigated in 165 CCA tissues. The effect of TNF-α on EMT-related CCA cell migration was evaluated using qRT-PCR, immunofluorescence and transwell migration assays.
  
  
  RESULTS
  ZEB2 and S100A4 mRNA levels were found to be higher in CCA tissues. High levels of S100A4 mRNA and ZEB2 protein were significantly associated with CCA metastasis (P = 0.04 and P = 0.03). Moreover, a trend toward statistical association was found with high levels of both ZEB2 mRNA and protein with shorter survival time (P = 0.10 and P = 0.19). In addition, TNF-α induced CCA cell migration by the induction of transforming growth factor-β (TGF-β) resulting in ZEB2 and S100A4 mRNA and protein activation.
  
  
  CONCLUSIONS
  These studies demonstrate that TNF-α plays crucial role in the progression of CCA by activating TGF-β signaling and the induction of ZEB2 and S100A4, EMT-related proteins expression.
authors.affiliation: Center for Research and Development of Medical Diagnostic Laboratories, Faculty of Associated Medical Sciences, Khon Kaen University, Khon Kaen, Thailand; Liver Fluke and Cholangiocarcinoma Research Center, Khon Kaen University, Khon Kaen, Thailand.
authors.forename: Anchalee
authors.initials: A
authors.lastname: Techasen
authors.forename: Nisana
authors.initials: N
authors.lastname: Namwat
authors.forename: Watcharin
authors.initials: W
authors.lastname: Loilome
authors.forename: Kassaporn
authors.initials: K
authors.lastname: Duangkumpha
authors.forename: Anucha
authors.initials: A
authors.lastname: Puapairoj
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Saya
authors.forename: Puangrat
authors.initials: P
authors.lastname: Yongvanit
mesh_terms: D000328:Adult; D000368:Aged; D014408:Biomarkers, Tumor / Q000378:metabolism; D045744:Cell Line, Tumor; D002465:Cell Movement; D018281:Cholangiocarcinoma / Q000378:metabolism* / Q000473:pathology*; D018450:Disease Progression; D005260:Female; D018398:Homeodomain Proteins / Q000378:metabolism*; D006801:Humans; D007124:Immunoenzyme Techniques; D008297:Male; D008856:Microscopy, Fluorescence; D008875:Middle Aged; D012333:RNA, Messenger / Q000378:metabolism*; D012097:Repressor Proteins / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000071999:S100 Calcium-Binding Protein A4; D009418:S100 Proteins / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000502:physiology*; D000074165:Zinc Finger E-box Binding Homeobox 2; D000071799:Zinc Finger E-box-Binding Homeobox 1
chemical_list: D014408:Biomarkers, Tumor; D018398:Homeodomain Proteins; D012333:RNA, Messenger; D012097:Repressor Proteins; D000071999:S100 Calcium-Binding Protein A4; D009418:S100 Proteins; D014157:Transcription Factors; D014409:Tumor Necrosis Factor-alpha; C486307:ZEB1 protein, human; C492363:ZEB2 protein, human; D000074165:Zinc Finger E-box Binding Homeobox 2; D000071799:Zinc Finger E-box-Binding Homeobox 1; C077591:S100A4 protein, human
keywords: Cholangiocarcinoma · Epithelial mesenchymal transition · S100A4 · ZEB2