Suppression of the Epidermal Growth Factor Receptor Inhibits Epithelial–Mesenchymal Transition in Human Pancreatic Cancer PANC-1 Cells

• Published in: Digestive diseases and sciences 2012-05, Vol.57 (5), p.1181-1189
• Main Authors: Chang, Zhi-Gang, Wei, Jun-Min, Qin, Chang-Fu, Hao, Kun, Tian, Xiao-Dong, Xie, Kun, Xie, Xue-Hai, Yang, Yin-Mo
• Format: Article
• Language: English
• Subjects: Analysis; Biochemistry; Cadherins - metabolism; Cancer; Cell Line, Tumor; Development and progression; Down-Regulation; Epidermal growth factor; Epithelial-Mesenchymal Transition - genetics; Fibronectins; Fibronectins - metabolism; Gastroenterology; Gene Knockdown Techniques - methods; Genetic Vectors; Hepatology; Humans; Lentivirus; Medicine; Medicine & Public Health; Oncology; Oncology, Experimental; Original Article; Pancreatic cancer; Pancreatic Neoplasms - genetics; Pancreatic Neoplasms - metabolism; Real-Time Polymerase Chain Reaction; Receptor, Epidermal Growth Factor - genetics; RNA; RNA, Small Interfering - genetics; RNA, Small Interfering - metabolism; Stem cells; Transcription Factors - metabolism; Transfection; Transplant Surgery; Vimentin - metabolism
• ISSN: 0163-2116; 1573-2568
• DOI: 10.1007/s10620-012-2036-4

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. Conclusion 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.