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Suppression of the Epidermal Growth Factor-like Domain 7 and Inhibition of Migration and Epithelial-Mesenchymal Transition in Human Pancreatic Cancer PANC-1 Cells

  • Wang, Yun-Liang (Department of General Surgery, the First Affiliated Hospital of Soochow University) ;
  • Dong, Feng-Lin (Department of Ultrasonography, the First Affiliated Hospital of Soochow University) ;
  • Yang, Jian (Department of General Surgery, the First Affiliated Hospital of Soochow University) ;
  • Li, Zhi (Department of Interventional Radiology, the First Affiliated Hospital of Soochow University) ;
  • Zhi, Qiao-Ming (Department of General Surgery, the First Affiliated Hospital of Soochow University) ;
  • Zhao, Xin (Department of General Surgery, the First Affiliated Hospital of Soochow University) ;
  • Yang, Yong (Department of General Surgery, the First Affiliated Hospital of Soochow University) ;
  • Li, De-Chun (Department of General Surgery, the First Affiliated Hospital of Soochow University) ;
  • Shen, Xiao-Chun (Department of Respiratory Medicine, the First Affiliated Hospital of Soochow University) ;
  • Zhou, Jin (Department of General Surgery, the First Affiliated Hospital of Soochow University)
  • Published : 2015.05.18

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. Materials and 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, NCPANC-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.

Keywords

References

  1. Batlle E, Sancho E, Franci C, et al (2000). The transcription factor snail is a repressor of E-cadherin gene expression in epithelial tumour cells. Nat Cell Biol, 2, 84-9. https://doi.org/10.1038/35000034
  2. Beuran M, Negoi I, Paun S, et al (2015). The epithelial to mesenchymal transition in pancreatic cancer: A systematic review. Pancreatology, [Epub ahead of print].
  3. Chang ZG, Wei JM, Qin CF, et al (2012). Suppression of the epidermal growth factor receptor inhibits epithelial-mesenchymal transition in human pancreatic cancer PANC-1 cells. Dig Dis Sci, 57, 1181-9. https://doi.org/10.1007/s10620-012-2036-4
  4. Chen WQ, Liang D, Zhang SW, et al (2013). Pancreatic cancer incidence and mortality patterns in china, 2009. Asian Pac J Cancer Prev, 14, 7321-4. https://doi.org/10.7314/APJCP.2013.14.12.7321
  5. Diaz R, Silva J, Garcia JM, et al (2008). Deregulated expression of miR-106a predicts survival in human colon cancer patients. Genes Chromosomes Cancer, 47, 794-802. https://doi.org/10.1002/gcc.20580
  6. Fan C, Yang LY, Wu F, et al (2013). The expression of Egfl7 in human normal tissues and epithelial tumors. Int J Biol Markers, 28, 71-83. https://doi.org/10.5301/JBM.2013.10568
  7. Fitch MJ, Campagnolo L, Kuhnert F, et al (2004). Egfl7, a novel epidermal growth factor-domain gene expressed in endothelial cells. Dev Dyn, 230, 316-24. https://doi.org/10.1002/dvdy.20063
  8. Hackert T, Buchler MW (2013). Pancreatic cancer: advances in treatment, results and limitations. Dig Dis, 31, 51-6. https://doi.org/10.1159/000347178
  9. Huang CH, Li XJ, Zhou YZ, et al (2010). Expression and clinical significance of EGFL7 in malignant glioma. J Cancer Res Clin Oncol, 136, 1737-43. https://doi.org/10.1007/s00432-010-0832-9
  10. Kurahara H, Takao S, Maemura K, et al (2012). Epithelial-mesenchymal transition and mesenchymal-epithelial transition via regulation of ZEB-1 and ZEB-2 expression in pancreatic cancer. J Surg Oncol, 105, 655-61. https://doi.org/10.1002/jso.23020
  11. Li Z, Ni CF, Zhou J, et al (2015). Expression of epidermal growth factor-like domain 7 is increased by transcatheter arterial embolization of liver tumors. Asian Pac J Cancer Prev, 16, 1191-6. https://doi.org/10.7314/APJCP.2015.16.3.1191
  12. Liu Y, Zhou YD, Xiao YL, et al (2015). Cyr61/CCN1 Overexpression Induces Epithelial-Mesenchymal Transition Leading to Laryngeal Tumor Invasion and Metastasis and Poor Prognosis. Asian Pac J Cancer Prev, 16, 2659-64. https://doi.org/10.7314/APJCP.2015.16.7.2659
  13. Liu SZ, Chen WQ, Wang N, et al(2014).Dietary factors and risk of pancreatic cancer: a multi-centre case-control study in China. Asian Pac J Cancer Prev, 15, 7947-50. https://doi.org/10.7314/APJCP.2014.15.18.7947
  14. Luo BH, Xiong F, Wang JP, et al (2014). Epidermal growth factor-like domain-containing protein 7 (EGFL7) enhances EGF receptor-AKT signaling, epithelial-mesenchymal transition, and metastasis of gastric cancer cells. PLoS One, 9, 99922. https://doi.org/10.1371/journal.pone.0099922
  15. Nichol D, Stuhlmann H (2012). EGFL7: a unique angiogenic signaling factor in vascular development and disease. Blood, 119, 1345-52. https://doi.org/10.1182/blood-2011-10-322446
  16. Parker LH, Schmidt M, Jin SW, et al (2004). The endothelial-cell-derived secreted factor Egfl7 regulates vascular tube formation. Nature, 428, 754-8. https://doi.org/10.1038/nature02416
  17. Soncin F, Mattot V, Lionneton F, et al (2003). VE-statin, an endothelial repressor of smooth muscle cell migration. Embo J, 22, 5700-11. https://doi.org/10.1093/emboj/cdg549
  18. Techasen A, Loilome W, Namwat N, et al (2012). Cytokines released from activated human macrophages induce epithelial mesenchymal transition markers of cholangiocarcinoma cells. Asian Pac J Cancer Prev, 13, 115-8.
  19. Thiery JP, Acloque H, Huang RYJ, et al (2009). Epithelial-Mesenchymal Transitions in Development and Disease. Cell, 139, 871-90. https://doi.org/10.1016/j.cell.2009.11.007
  20. Wu F, Yang LY, Li YF, et al (2009). Novel role for epidermal growth factor-like domain 7 in metastasis of human hepatocellular carcinoma. Hepatol, 50, 1839-50. https://doi.org/10.1002/hep.23197
  21. Xu HF, Chen L, Liu XD, et al (2014). Targeting EGFL7 expression through RNA interference suppresses renal cell carcinoma growth by inhibiting angiogenesis. Asian Pac J Cancer Prev, 15, 3045-50. https://doi.org/10.7314/APJCP.2014.15.7.3045
  22. Zhou L, Li J, Zhao Y, et al (2014). Prognostic significance of epidermal growth factor-like domain 7 in pancreatic cancer. Hepatobiliary Pancreat Dis Int, 13, 523-8. https://doi.org/10.1016/S1499-3872(14)60272-1

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