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Suppression of the Epidermal Growth Factor-like Domain 7 and Inhibition of Migration and Epithelial-Mesenchymal Transition in Human Pancreatic Cancer PANC-1 Cells

  • Wang, Yun-Liang (Department of General Surgery, the First Affiliated Hospital of Soochow University) ;
  • Dong, Feng-Lin (Department of Ultrasonography, the First Affiliated Hospital of Soochow University) ;
  • Yang, Jian (Department of General Surgery, the First Affiliated Hospital of Soochow University) ;
  • Li, Zhi (Department of Interventional Radiology, the First Affiliated Hospital of Soochow University) ;
  • Zhi, Qiao-Ming (Department of General Surgery, the First Affiliated Hospital of Soochow University) ;
  • Zhao, Xin (Department of General Surgery, the First Affiliated Hospital of Soochow University) ;
  • Yang, Yong (Department of General Surgery, the First Affiliated Hospital of Soochow University) ;
  • Li, De-Chun (Department of General Surgery, the First Affiliated Hospital of Soochow University) ;
  • Shen, Xiao-Chun (Department of Respiratory Medicine, the First Affiliated Hospital of Soochow University) ;
  • Zhou, Jin (Department of General Surgery, the First Affiliated Hospital of Soochow University)
  • Published : 2015.05.18

Abstract

Background: Epidermal growth factor-like domain multiple 7 (EGFL7), a secreted protein specifically expressed by endothelial cells during embryogenesis, recently was identified as a critical gene in tumor metastasis. Epithelial-mesenchymal transition (EMT) was found to be closely related with tumor progression. Accordingly, it is important to investigate the migration and EMT change after knock-down of EGFL7 gene expression in human pancreatic cancer cells. Materials and Methods: EGFL7 expression was firstly testified in 4 pancreatic cancer cell lines by real-time polymerase chain reaction (Real-time PCR) and western blot, and the highest expression of EGFL7 was found in PANC-1 cell line. Then, PANC-1 cells transfected with small interference RNA (siRNA) of EGFL7 using plasmid vector were named si-PANC-1, while transfected with negative control plasmid vector were called NC-PANC-1. Transwell assay was used to analyze the migration of PANC-1 cells. Real-time PCR and western blotting were used to detect the expression change of EGFL7 gene, EMT markers like E-Cadherin, N-Cadherin, Vimentin, Fibronectin and transcription factors like snail, slug in PANC-1, NCPANC-1, and si-PANC-1 cells, respectively. Results: After successful plasmid transfection, EGFL7 gene were dramatically knock-down by RNA interference in si-PANC-1 group. Meanwhile, migration ability decreased significantly, compared with PANC-1 and NC-PANC-1 group. Meanwhile, the expression of epithelial phenotype marker E-Cadherin increased and that of mesenchymal phenotype markers N-Cadherin, Vimentin, Fibronectin dramatically decreased in si-PANC-1 group, indicating a reversion of EMT. Also, transcription factors snail and slug decreased significantly after RNA interference. Conclusions: Current study suggested that highly-expressed EGFL7 promotes migration of PANC-1 cells and acts through transcription factors snail and slug to induce EMT, and further study is needed to confirm this issue.

Keywords

Pancreatic cancer;epidermal growth factor-like domain multiple 7;epithelial-mesenchymal transition

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