Suppression of the Epidermal Growth Factor Receptor Inhibits Epithelial–Mesenchymal Transition in Human Pancreatic Cancer PANC-1 Cells
- 641 Downloads
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.
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.
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.
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.
KeywordsPancreatic cancer Epidermal growth factor receptor Epithelial–mesenchymal transition E-cadherin
Epidermal growth factor receptor
Epidermal growth factor
Dulbecco’s modified Eagle’s medium
Fetal bovine serum
Bovine serum albumin
Sodium dodecyl sulfate polyacrylamide gel electrophoresis
Tyrosine kinase inhibitor
Small interfering RNA
This work was supported by the National Basic Research Program Grant (no. 30972897) and the National Natural Science Foundation, People’s Republic of China.
Conflict of interest
No conflicts of interest have been declared by all authors.
- 2.Landis SH, Murray T, Bolden S, et al. Cancer statistics, 1999. CA Cancer J Clin. 1999;49:8–31.Google Scholar
- 4.Wang Y, Zhou BP. Epithelial–mesenchymal transition in breast cancer progression and metastasis. Chin J Cancer. 2011;30:603–611.Google Scholar
- 15.Li Y, VandenBoom TG, Kong D, et al. Up-regulation of miR-200 and let-7 by natural agents leads to the reversal of epithelial-to-mesenchymal transition in gemcitabine-resistant pancreatic cancer cells. Cancer Res 2009;69:6704–6712.Google Scholar
- 16.Wang Z, Li Y, Ahmad A, et al. Pancreatic cancer: understanding and overcoming chemoresistance. Nat Rev Gastroenterol Hepatol. 2011;8:27–33.Google Scholar
- 20.Al Moustafa AE, Yen L, Benlimame N, et al. Regulation of E-cadherin/catenin complex patterns by epidermal growth factor receptor modulation in human lung cancer cells. Lung Cancer. 2002;37:49–56.Google Scholar
- 37.Mimeault M, Johansson SL, Vankatraman G, et al. Combined targeting of epidermal growth factor receptor and hedgehog signaling by gefitinib and cyclopamine cooperatively improves the cytotoxic effects of docetaxel on metastatic prostate cancer cells. Mol Cancer Ther. 2007;6:967–978.PubMedCrossRefGoogle Scholar