Skip to main content

Advertisement

SpringerLink
Log in

Pancreatic tumor growth is regulated by the balance between positive and negative modulators of angiogenesis

  • Published:
Angiogenesis Aims and scope Submit manuscript

Abstract

There is increasing evidence for the implication of tumor-derived angiogenic and anti-angiogenic factors in controlling tumor growth in vivo. In this study, we documented the production of inhibitors of angiogenesis by pancreatic cancer cells and examined how changes in the balance between pro- and anti-angiogenic factors regulate tumor growth in vivo. The human pancreatic cancer cell line Hs-776T (HS-W) produces slow-growing tumors in SCID mice. Cells of a variant form (HS-R) of Hs-776T produced faster-growing tumors compared to HS-W. Characterization of HS-W and HS-R cells in vitro showed similar proliferation rates and production of the angiogenic factors vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF). Analyzes of anti-angiogenic factors showed comparable levels of angiostatin and thrombospondin 1 and 2, but endostatin was only detected in conditioned media of HS-W cells and was absent in HS-R. Cell proliferation was similar in both tumor types in vivo, whereas HS-W tumors demonstrated increased apoptosis with a high percentage of apoptotic endothelial cells (EC). Subsequently, VEGF was over-expressed in Hs-776T cells (HS-VF), resulting in rapidly growing tumors and lowering tumor and EC apoptosis. Collectively, our study confirms that tumor growth is dependent on its ability to increase the angiogenic stimulus or to reduce the amounts of endogenous anti-angiogenic factors.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Risau W. Mechanisms of angiogenesis. Nature 1997; 386: 671–4.

    Google Scholar 

  2. Folkman J. Tumor angiogenesis: Therapeutic implications. N Engl J Med 1971; 285: 1182–6.

    Google Scholar 

  3. Plate KH, Breier G, Weich HA et al. Vascular endothelial growth factor is a potential tumor angiogenesis factor in gliomas in vivo. Nature 1992; 359: 845–8.

    Google Scholar 

  4. Takahashi Y, Kitadai Y, Bucana CD et al. Expression of vascular endothelial growth factor and its receptor, KDR, correlates with vascularity, metastasis and proliferation in human colon cancer. Cancer Res 1995; 55:3964–8.

    Google Scholar 

  5. Zhang HAT, Craft P, Scott PAE et al. Enhancement of tumor growth and vascular density by transfection of vascular endothelial growth factor into MCF-7 human breast carcinoma cells. J Natl Cancer Inst 1995; 87: 213–8.

    Google Scholar 

  6. Belletti B, Ferraro P, Arra C et al. Modulation of in vivo growth of thyroid tumor-derived cell lines by sense and antisense vascular endothelial growth factor gene. Oncogene 1999; 18: 4860–9.

    Google Scholar 

  7. Folkman J. What is the evidence that tumors are angiogenesis dependent? J Natl Cancer Inst 1990; 82: 4–6.

    Google Scholar 

  8. Dirix LY, Vermeulen PB, Pawinski A et al. Elevated levels of the angiogenic cytokines basic fibroblast growth factor and vascular endothelial growth factor in sera of cancer patients. Br J Cancer 1997; 76: 238–43.

    Google Scholar 

  9. Gasparini G, Toi M, Gion M et al. Prognostic significance of vascular endothelial growth factor protein in node-negative breast carcinoma. J Natl Cancer Inst 1997; 89: 139–47.

    Google Scholar 

  10. Linderholm B, Lindh B, Tavelin B et al. P53 and vascular-endothelial-growth-factor (VEGF) expression predicts outcome in 833 patients with primary breast carcinoma. Int J Cancer 2000; 89: 51–62.

    Google Scholar 

  11. Ikeda N, Adachi M, Taki T et al. Prognostic significance of angiogenesis in human pancreatic cancer. Br J Cancer 1999; 79: 1553–63.

    Google Scholar 

  12. Takeda A, Shimada H, Imaseki H et al. Clinical significance of serum vascular endothelial growth factor in colorectal cancer patients: Correlation with clinicopathological factors and tumor markers. Oncol Rep 2000; 7: 333–8.

    Google Scholar 

  13. Sauter ER, Nesbit M, Watson JC et al. Vascular endothelial growth factor is a marker of tumor invasion and metastasis in squamous cell carcinomas of the head and neck. Clin Cancer Res 1999; 5: 775–82.

    Google Scholar 

  14. Duque JL, Loughlin KR, Adam RM et al. Plasma levels of vascular endothelial growth factor are increased in patients with metastatic prostate cancer. Urology 1999; 4: 523–7.

    Google Scholar 

  15. Aguayo A, Estey E, Kantarjian H et al. Cellular vascular endothelial growth factor is a predictor of outcome in patients with acute myeloid leukemia. Blood 1999; 94: 3717–21.

    Google Scholar 

  16. Ribatti D, Vacca A, Nico B et al. Bone marrow angiogenesis and mast cell density increase simultaneously with progression of human multiple myeloma. Br J Cancer 1999; 79: 451–5.

    Google Scholar 

  17. Salven P, Teerenhovi L, Joensuu H. A high pretreatment serum basic fibroblast growth factor concentration is an independent predictor of poor prognosis in non-Hodgkin's lymphoma. Blood 1999; 94: 3334–9.

    Google Scholar 

  18. O'Reilly MS, Holmgren L, Shing Y et al. Angiostatin: A novel angiogenesis inhibitor that mediates the suppression of metastases by a Lewis lung carcinoma. Cell 1994; 79: 315–28.

    Google Scholar 

  19. Folkman J. Angiogenesis inhibitors generated by tumors. Mol Med 1995; 1: 120–2.

    Google Scholar 

  20. O'Reilly MS, Boehm T, Shing Y et al. Endostatin: An endogenous inhibitor of angiogenesis and tumor growth. Cell 1997; 88: 277–85.

    Google Scholar 

  21. Cao Y, Ji RW, Davidson D et al. Kringle domains of human angiostatin. Characterization of the anti-proliferative activity on endothelial cells. J Biol Chem 1996; 271: 29461–7.

    Google Scholar 

  22. Dhanabal M, Ramchandran R, Waterman MJ et al. Endostatin induces endothelial cell apoptosis. J Biol Chem 1999; 274: 11721–6.

    Google Scholar 

  23. Boehm T, Folkman J, Browder T et al. Antiangiogenic therapy of experimental cancer does not induce acquired drug resistance. Nature 1997; 390: 404–7.

    Google Scholar 

  24. Bornstein P. Thrombospondins: Structure and regulation of expression. FASEB J 1992; 6: 3290–9.

    Google Scholar 

  25. Streit M, Riccardi L, Velasco P et al. Thrombospondin-2: A potent endogenous inhibitor of tumor growth and angiogenesis. Proc Natl Acad Sci USA 1999; 96: 14888–93.

    Google Scholar 

  26. Cao Y, O'Reilly MS, Marshall B et al. Expression of angiostatin cDNA in a murine fibrosarcoma suppresses primary tumor growth and produces long-term dormancy of metastases. J Clin Invest 1998; 101: 1055–63.

    Google Scholar 

  27. Chen QR, Kumar D, Stass SA et al. Liposomes complexed to plasmids encoding angiostatin and endostatin inhibit breast cancer in nude mice. Cancer Res 1999; 59: 3308–12.

    Google Scholar 

  28. Blezinger P, Wang J, Gondo M et al. Systemic inhibition of tumor growth and tumor metastases by intramuscular administration of the endostatin gene. Nat Biotechnol 1999; 17: 343–8.

    Google Scholar 

  29. Atlas of Cancer Mortality in the United States 1950–1994. National Cancer Institute 1999. Bethesda, Maryland: National Institute of Health.

  30. Warshaw AL, Fernandez-del Castillo C. Pancreatic carcinoma. N Engl J Med 1992; 326: 455–65.

    Google Scholar 

  31. Kuehn R, Lelkes PI, Bloechle C et al. Angiogenesis, angiogenic growth factors, and cell adhesion molecules are upregulated in chronic pancreatic diseases: Angiogenesis in chronic pancreatitis and in pancreatic cancer. Pancreas 1999; 18: 96–103.

    Google Scholar 

  32. Ellis LM, Takahashi Y, Fenoglio CJ et al. Vessel counts and vascular endothelial growth factor expression in pancreatic adenocarcinoma. Eur J Cancer 1998; 34: 337–40.

    Google Scholar 

  33. Fujimoto K, Hosotani R, Wada M et al. Expression of two angiogenic factors, vascular endothelial growth factor and plateletderived endothelial cell growth factor in human pancreatic cancer, and its relationship to angiogenesis. Eur J Cancer 1998; 34: 1439–47.

    Google Scholar 

  34. Itakura J, Ishiwata T, Friess H et al. Enhanced expression of vascular endothelial growth factor in human pancreatic cancer correlates with local disease progression. Clin Cancer Res 1997; 3: 1309–16.

    Google Scholar 

  35. Knoll MR, Rudnitzki D, Sturm J et al. Correlation of postoperative survival and angiogenic growth factors in pancreatic carcinoma. Hepatogastroenterology 2001; 48: 1162–5.

    Google Scholar 

  36. Stipa F, Lucandri G, Limiti MR et al. Angiogenesis as a prognostic indicator in pancreatic ductal adenocarcinoma. Anticancer Res 2002; 22: 445–9.

    Google Scholar 

  37. Shishido T, Yasoshima T, Denno R et al. Inhibition of liver metastasis of human pancreatic carcinoma by angiogenesis inhibitor TNP-470in combination with cisplatin. Jpn J Cancer Res 1998; 89: 963–9.

    Google Scholar 

  38. Kisker O, Onizuka S, Banyard J et al. Generation of multiple angiogenesis inhibitors by human pancreatic cancer. Cancer Res 2001; 61: 7298–304.

    Google Scholar 

  39. Hruban RH, Wilentz RE, Kern SE. Genetic progression in the pancreatic ducts. Am J Pathol 2000; 156: 1821–5.

    Google Scholar 

  40. Bignami M, Rosa S, Falcone G et al. Specific viral oncogenes cause differential effects on cell-to-cell communication, relevant to the suppression of the transformed phenotype by normal cells. Mol Carcinog 1988; 1: 67–75.

    Google Scholar 

  41. Oshika Y, Masuda K, Tokunaga T et al. Thrombospondin 2 gene expression is correlated with decreased vascularity in non-small cell lung cancer. Clin Cancer Res 1998; 4: 1785–8.

    Google Scholar 

  42. Varani J, Riser BL, Hughes LA et al. Characterization of thrombospondin synthesis, secretion and cell surface expression by human tumor cells. Clin Exp Metastasis 1989; 7: 265–76.

    Google Scholar 

  43. Kondo Y, Arii S, Mori A et al. Enhancement of angiogenesis, tumor growth, and metastasis by transfection of vascular endothelial growth factor into LoVo human colon cancer cell line. Clin Cancer Res 2000; 6: 622–30.

    Google Scholar 

  44. Shi YP, Ferrara N. Oncogenic ras fails to restore an in vivo tumorigenic phenotype in embryonic stem cells lacking vascular endothelial growth factor (VEGF). Biochem Biophys Res Commun 1999; 254: 480–3.

    Google Scholar 

  45. Ferrara N. Molecular and biological properties of vascular endothelial growth factor. J Mol Med 1999; 77: 527–43.

    Google Scholar 

  46. Itakura J, Ishiwata T, Shen B et al. Concomitant over-expression of vascular endothelial growth factor and its receptors in pancreatic cancer. Int J Cancer 2000; 85: 27–34.

    Google Scholar 

  47. Graeven U, Fiedler W, Karpinski S et al. Melanoma-associated expression of vascular endothelial growth factor and its receptors FLT-1 and KDR. J Cancer Res Clin Oncol 1999; 125: 621–9.

    Google Scholar 

  48. Dvorak HF, Sioussat TM, Brown LF et al. Distribution of vascular endothelial growth factor in tumors: Concentration in tumor blood vessels. J Exp Med 1991; 174: 1275–8.

    Google Scholar 

  49. Springer ML, Chen AS, Kraft PE et al. VEGF gene delivery to muscle: Potential role for vasculogenesis in adults. Mol Cell 1998; 2: 549–58.

    Google Scholar 

  50. Lee RJ, Springer ML, Blanco-Bose WE et al. VEGF gene delivery to myocardium: Deleterious effects of unregulated expression. Circulation 2000; 102: 898–901.

    Google Scholar 

  51. Miller JW, Adamis AP, Aiello LP. Vascular endothelial growth factor in ocular neovascularization and proliferative diabetic retinopathy. Diabetes Metab Rev 1997; 13: 37–50.

    Google Scholar 

  52. Keck PJ, Hauser SD, Krivi G et al. Vascular permeability factor, an endothelial cell mitogen related to PDGF. Science 1989; 246: 1309–12.

    Google Scholar 

  53. Midy V, Plouet J. Vasculotropin/vascular endothelial growth factor induces differentiation in cultured osteoblasts. Biochem Biophys Res Commun 1994; 199: 380–6.

    Google Scholar 

  54. von Marschall Z, Cramer T, Hocker M et al. De novo expression of vascular endothelial growth factor in human pancreatic cancer: Evidence for an autocrine mitogenic loop. Gastroenterology 2000; 119: 1358–72.

    Google Scholar 

  55. Bunone G, Vigneri P, Mariani L et al. Expression of angiogenesis stimulators and inhibitors in human thyroid tumors and correlation with clinical pathological features. Am J Pathol 1999; 155: 1967–76.

    Google Scholar 

  56. Baker CH, Solorzano CC, Fidler IJ. Blockade of vascular endothelial growth factor receptor and epidermal growth factor receptor signaling for therapy of metastatic human pancreatic cancer. Cancer Res 2002; 62: 1996–2003.

    Google Scholar 

  57. Shishido T, Yasoshima T, Denno R et al. Inhibition of liver metastasis of human pancreatic carcinoma by angiogenesis inhibitor TNP-470in combination with cisplatin. Jpn J Cancer Res 1998; 89: 963–9.

    Google Scholar 

  58. Bruns CJ, Solorzano CC, Harbison MT et al. Blockade of the epidermal growth factor receptor signaling by a novel tyrosine kinase inhibitor leads to apoptosis of endothelial cells and therapy of human pancreatic carcinoma. Cancer Res 2000; 60: 2926–35.

    Google Scholar 

  59. Yanagi K, Onda M, Uchida. Effect of angiostatin on liver metastasis of pancreatic cancer in hamsters. Jpn J Cancer Res 2000; 91: 723–30.

    Google Scholar 

  60. Kisker O, Becker CM, Prox D et al. Continuous administration of endostatin by intraperitoneally implanted osmotic pump improves the efficacy and potency of therapy in a mouse xenograft tumor model. Cancer Res 2001; 6: 7669–74.

    Google Scholar 

  61. Indraccolo S, Gola E, Rosato A et al. Differential effects of angiostatin, endostatin and interferon-alpha(1) gene transfer on in vivo growth of human breast cancer cells. Gene Ther 2002; 9: 867–78.

    Google Scholar 

  62. Ziche M, Sorriso C, Parenti A et al. Biological parameters for the choice of antiangiogenic therapy and efficacy monitoring. Int J Biol Markers 1999; 14: 214–7.

    Google Scholar 

Download references

Author information

Author notes

  1. Gunter Schuch

    Present address: Department of Oncology and Hematology, University Hospital Hamburg-Eppendorf, Hamburg, Germany

Authors and Affiliations

  1. Department of Urology, The Laboratory for Cellular Therapeutics and Tissue Engineering, Children's Hospital, Harvard Medical School, Boston, Massachusetts, USA

    Gunter Schuch, Anthony Atala & Shay Soker

  2. The Laboratory for Surgical Research, Children's Hospital, Harvard Medical School, Boston, Massachusetts, USA

    Oliver Kisker

Authors
  1. Gunter Schuch
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Oliver Kisker
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Anthony Atala
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Shay Soker
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Shay Soker.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Schuch, G., Kisker, O., Atala, A. et al. Pancreatic tumor growth is regulated by the balance between positive and negative modulators of angiogenesis. Angiogenesis 5, 181–190 (2002). https://doi.org/10.1023/A:1023893931057

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1023893931057

Search

Navigation