title: Suppression of the Epidermal Growth Factor-like Domain 7 and Inhibition of Migration and Epithelial-Mesenchymal Transition in Human Pancreatic Cancer PANC-1 Cells.
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.
  
  
  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, NC- PANC-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.
authors.affiliation: Department of General Surgery, the First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China E-mail : 13913500369@163.com; sxcsmile20008@163.com.
authors.forename: Yun-Liang
authors.initials: YL
authors.lastname: Wang
authors.forename: Feng-Lin
authors.initials: FL
authors.lastname: Dong
authors.forename: Jian
authors.initials: J
authors.lastname: Yang
authors.forename: Zhi
authors.initials: Z
authors.lastname: Li
authors.forename: Qiao-Ming
authors.initials: QM
authors.lastname: Zhi
authors.forename: Xin
authors.initials: X
authors.lastname: Zhao
authors.forename: Yong
authors.initials: Y
authors.lastname: Yang
authors.forename: De-Chun
authors.initials: DC
authors.lastname: Li
authors.forename: Xiao-Chun
authors.initials: XC
authors.lastname: Shen
authors.forename: Jin
authors.initials: J
authors.lastname: Zhou
mesh_terms: D017209:Apoptosis; D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism; D015153:Blotting, Western; D002135:Calcium-Binding Proteins; D002465:Cell Movement; D049109:Cell Proliferation; D066255:EGF Family of Proteins; D016228:Endothelial Growth Factors / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D058750:Epithelial-Mesenchymal Transition; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D010190:Pancreatic Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology*; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D014407:Tumor Cells, Cultured
chemical_list: D014408:Biomarkers, Tumor; D002135:Calcium-Binding Proteins; D066255:EGF Family of Proteins; C527877:EGFL7 protein, human; D016228:Endothelial Growth Factors; D012333:RNA, Messenger; D034741:RNA, Small Interfering