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Annals of Surgical Oncology

, Volume 19, Issue 8, pp 2733–2743 | Cite as

VEGF-A/VEGFR-2 Signaling Plays an Important Role for the Motility of Pancreas Cancer Cells

  • Yosuke Doi
  • Masakazu YashiroEmail author
  • Nobuya Yamada
  • Ryosuke Amano
  • Satoru Noda
  • Kosei Hirakawa
Translational Research and Biomarkers

Abstract

Background

Pancreatic cancer is one of the most lethal solid tumors. Vascular endothelial growth factor receptors (VEGFRs) are expressed not only by endothelial cells but also by pancreatic cancer cells. VEGFRs might play an important role for the development of pancreatic cancer cells. The purpose of this study was to evaluate the efficacy of VEGF/VEGFR-2—targeted therapy in pancreatic carcinoma.

Methods

Five pancreatic carcinoma cell lines were used. The expression level of VEGFR-2 of cancer cells was examined by RT-PCR and Western blot. The effects of VEGFs, bevacizumab as an anti-VEGF antibody, sunitinib as a tyrosine kinase inhibitor against VEGFRs, and VEGF-R2 siRNA on the motility activity of pancreatic cancer cells were examined by invasion assay and wound healing assay. The effect of VEGF, bevacizumab, and sunitinib on the phosphorylation of VEGFR-2 and downstream effecter molecules, MAPK and PI3K, was examined by western blot.

Results

Pancreatic cancer cell lines expressed VEGFR-2. VEGF-A significantly increased the motility of pancreas cancer cells, which was inhibited by VEGFR-2 siRNA. Conditioned medium from pancreas cancer cells significantly stimulated the motility of pancreas cancer cells. VEGF/VEGFR inhibitors, bevacizumab and sunitinib, significantly decreased the motility of pancreas cancer cells. VEGFR-2 phosphorylation level of pancreas cancer cells was increased by VEGF-A. Bevacizumab and sunitinib decreased the level of VEGFR-2 phosphorylation, p-ERK, and p-Akt expression. VEGF-A decreased zonula occludens (ZO-1) or ZO-2 expression in pancreas cancer cells.

Conclusions

VEGF-A/VEGFR-2 signaling plays an important role in inducing invasion and migration of pancreatic cancer cells.

Keywords

Vascular Endothelial Growth Factor Pancreatic Cancer Bevacizumab Conditioned Medium Sunitinib 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgment

This work was supported by KAKENHI (Grant-in-Aid for Scientific Research, No. 18591475, 20591073, 18390369, 22390262, and 23390329) and by a Grant-in Aid for the Sagawa Foundation for Cancer Research.

Conflict of interest

The authors declare that they have no conflicts of interest to disclose.

Supplementary material

Supplementary movie 1 Real-time living-cell imaging of MiaPaCa-2 cells was monitored by time-lapse video microscopy for 72 h at 15-min intervals. VEGF-A significantly increased the motility of MiaPaCa-2 cells by the wound healing assay (MOV 2,490 kb)

Supplementary movie 2 Real-time living-cell imaging of MiaPaCa-2 cells was monitored by time-lapse video microscopy for 72 h at 15-min intervals. Bevacizumab decreased the migrating activity of MiaPaCa-2 cells (MOV 2,400 kb)

10434_2011_2181_MOESM3_ESM.eps (770 kb)
Supplement Fig. A Effect of VEGFs on the proliferation of pancreas cancer cells. The proliferative activity of pancreas cancer cells was not affected by VEGF (EPS 771 kb)
10434_2011_2181_MOESM4_ESM.eps (781 kb)
Supplement Fig. B Effect of inhibitors on the proliferative activity of pancreas cancer cells. The mitogenic activity of pancreas cancer cells was not affected by inhibitors, bevacizumab, sunitinib, and lapatinib, examined (EPS 781 kb)
10434_2011_2181_MOESM5_ESM.eps (8.9 mb)
Supplement Figs. C Effect of VEGF-A and signal inhibitors on migration activity of pancreas cancer cells. C Representative phase-contrast images of the wound-healing assay. The number of migrating MiaPaCa-2 cells was increased by VEGF-A and decreased by bevacizumab and sunitinib. Dotted line, wound margin. (EPS 9,067 kb)
10434_2011_2181_MOESM6_ESM.eps (785 kb)
Supplement Figs. D Effect of VEGF-A and signal inhibitors on migration activity of pancreas cancer cells. VEGF stimulated the migration of pancreas cancer cells, MiaPaCa-2, RWP-1, and OCUP-AT. Bevacizumab and sunitinib significantly decreased the number of migratory MiaPaCa-2, RWP-1, and OCUP-AT cells but not the number of migratory Panc-1 and SW1990 cells. Lapatinib did not affect the migration of any pancreatic cells. Four randomly chosen fields were counted for each assay. The mean of four fields was calculated as the sample value. For each group, the culture was performed in triplicate. Data are presented as the mean ± SD (three samples per group). *p < 0.05 vs. control; **p < 0.01 vs. control (EPS 786 kb)
10434_2011_2181_MOESM7_ESM.eps (3.9 mb)
Supplement Fig. E The mRNA expression level of ZO-1 in MiaPaCa-2 and OCUP-AT with VEGF-A was lower than that in these cell lines without VEGF-A, and the mRNA expression level of ZO-2 in MiaPaCa-2 and RWP-1 with VEGF-A was lower than that in these cell lines without VEGF-A (EPS 4,017 kb)

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Copyright information

© Society of Surgical Oncology 2011

Authors and Affiliations

  • Yosuke Doi
    • 1
  • Masakazu Yashiro
    • 1
    • 2
    Email author
  • Nobuya Yamada
    • 1
  • Ryosuke Amano
    • 1
  • Satoru Noda
    • 1
  • Kosei Hirakawa
    • 1
  1. 1.Department of Surgical OncologyOsaka City University Graduate School of MedicineOsakaJapan
  2. 2.Oncology Institute of Geriatrics and Medical ScienceOsaka City University Graduate School of MedicineOsakaJapan

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