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Digestive Diseases and Sciences

, Volume 57, Issue 5, pp 1181–1189 | Cite as

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

  • Zhi-Gang Chang
  • Jun-Min Wei
  • Chang-Fu Qin
  • Kun Hao
  • Xiao-Dong Tian
  • Kun Xie
  • Xue-Hai Xie
  • Yin-Mo YangEmail author
Original Article

Abstract

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.

Keywords

Pancreatic cancer Epidermal growth factor receptor Epithelial–mesenchymal transition E-cadherin 

Abbreviations

EGFR

Epidermal growth factor receptor

EGF

Epidermal growth factor

EMT

Epithelial–mesenchymal transition

DMEM

Dulbecco’s modified Eagle’s medium

FBS

Fetal bovine serum

BSA

Bovine serum albumin

RIPA

Radioimmunoprecipitation assay

SDS-PAGE

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

HRP

Horseradish peroxidase

DAPI

4′,6-Diamidino-2-phenylindole

TKI

Tyrosine kinase inhibitor

RNAi

RNA interference

siRNA

Small interfering RNA

ECM

Extracellular matrix

Notes

Acknowledgments

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.

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Zhi-Gang Chang
    • 1
    • 2
  • Jun-Min Wei
    • 2
  • Chang-Fu Qin
    • 1
  • Kun Hao
    • 1
  • Xiao-Dong Tian
    • 1
  • Kun Xie
    • 1
  • Xue-Hai Xie
    • 1
  • Yin-Mo Yang
    • 1
    Email author
  1. 1.Department of General SurgeryPeking University First HospitalBeijingPeople’s Republic of China
  2. 2.Department of General SurgeryBeijing Hospital, Ministry of HealthBeijingPeople’s Republic of China

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