Abstract
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. 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. 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. 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.
Highlights
Pancreatic carcinoma is the 4th leading cause of cancerrelated death in the world (Chen et al, 2013; Liu et al, 2014; Siegel et al, 2015)
Materials and Methods: Epidermal growth factor-like domain multiple 7 (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
Zhou et al reported that EGFL7 expression in pancreatic carcinoma tissues was significantly higher than that in non-tumor tissues, and high expression of EGFL7 in pancreatic carcinoma tissues was significantly associated with poor overall survival, accompanied by several conventional clinicopathological variables, such
Summary
Pancreatic carcinoma is the 4th leading cause of cancerrelated death in the world (Chen et al, 2013; Liu et al, 2014; Siegel et al, 2015). Epidermal growth factor-like domain multiple 7 (EGFL7) is a secreted protein that contains two EGFlike domains and is conserved across species (Nichol and Stuhlmann, 2012) It was initially regarded as an endothelial cell-specific gene and an important regulator in tubulogenesis during embryonic development (Fitch et al, 2004). Epidermal growth factor-like domain multiple 7 (EGFL7), a secreted protein expressed by endothelial cells during embryogenesis, recently was identified as a critical gene in tumor metastasis. It is important to investigate the migration and EMT change after knock-down of EGFL7 gene expression in human pancreatic cancer 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. 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
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