title: Suppression of the epidermal growth factor receptor inhibits epithelial-mesenchymal transition in human pancreatic cancer PANC-1 cells.
abstract: BACKGROUND
  Aberrant expression of epidermal growth factor receptor (EGFR) has been detected in pancreatic cancer; however, the mechanisms of EGFR in inducing pancreatic cancer development have not been adequately elucidated. The objective of this study was to determine the role of EGFR in mediating epithelial-mesenchymal transition (EMT) in pancreatic cancer cells.
  
  
  METHODS
  Pancreatic cancer cell line PANC-1 was transfected with small interfering RNA of EGFR by use of a lentiviral expression vector to establish an EGFR-knockdown cell line (si-PANC-1). PANC-1 cells transfected with lentiviral vector expressing negative control sequence were used as negative control (NC-PANC-1). Scratch assay and transwell study were used to analyze cell migration and invasion. Real-time PCR and Western blotting were used to detect the expression of EMT markers E-cadherin, N-cadherin, vimentin, and fibronectin and transcription factors snail, slug, twist1, and sip1 in PANC-1, NC-PANC-1, and si-PANC-1 cells. Immunofluorescent staining with these antibodies and confocal microscopy were used to observe their cellular location and morphologic changes.
  
  
  RESULTS
  After RNA interference of EGFR, the migration and invasion ability of si-PANC-1 cells decreased significantly. The expression of epithelial phenotype marker E-cadherin increased and the expression of mesenchymal phenotype markers N-cadherin, vimentin, and fibronectin decreased, indicating reversion of EMT. We also observed intracellular translocation of E-cadherin. Expression of transcription factors snail and slug in si-PANC-1 cells decreased significantly.
  
  
  CONCLUSIONS
  Suppression of EGFR expression can significantly inhibit EMT of pancreatic cancer PANC-1 cells. The mechanism may be related with the down-regulation of the expression of transcription factors snail and slug.
authors.affiliation: Department of General Surgery, Peking University First Hospital, 8th Xishiku Street, Xicheng District, Beijing 100034, People's Republic of China.
authors.forename: Zhi-Gang
authors.initials: ZG
authors.lastname: Chang
authors.forename: Jun-Min
authors.initials: JM
authors.lastname: Wei
authors.forename: Chang-Fu
authors.initials: CF
authors.lastname: Qin
authors.forename: Kun
authors.initials: K
authors.lastname: Hao
authors.forename: Xiao-Dong
authors.initials: XD
authors.lastname: Tian
authors.forename: Kun
authors.initials: K
authors.lastname: Xie
authors.forename: Xue-Hai
authors.initials: XH
authors.lastname: Xie
authors.forename: Yin-Mo
authors.initials: YM
authors.lastname: Yang
mesh_terms: D015820:Cadherins / Q000378:metabolism; D045744:Cell Line, Tumor; D015536:Down-Regulation; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics*; D066246:ErbB Receptors / Q000235:genetics*; D005353:Fibronectins / Q000378:metabolism; D055785:Gene Knockdown Techniques / Q000379:methods; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus; D010190:Pancreatic Neoplasms / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D014157:Transcription Factors / Q000378:metabolism; D014162:Transfection; D014746:Vimentin / Q000378:metabolism
chemical_list: D015820:Cadherins; D005353:Fibronectins; D034741:RNA, Small Interfering; D014157:Transcription Factors; D014746:Vimentin; D066246:ErbB Receptors