Clinical & Experimental Metastasis

, Volume 21, Issue 2, pp 107–118 | Cite as

Vascular endothelial growth factor expression promotes the growth of breast cancer brain metastases in nude mice

  • Lee Su Kim
  • Suyung Huang
  • Weixin Lu
  • Dina Chelouche Lev
  • Janet E. Price
Article

Abstract

Patients with breast cancer brain metastases cannot be cured and have a poor prognosis, with a median survival time of six months after diagnosis, despite developments in diagnostic and therapeutic modalities. In large part the progress in understanding the biology of breast cancer brain metastasis has been limited by the lack of suitable cell lines and experimental models. The objective of this study was to develop a reliable experimental model to study the pathogenesis of breast cancer brain metastases, using intra-internal carotid artery injection of breast cancer cells into nude mice. Brain metastasis-selected variant cells were recovered after three cycles of injection into the internal carotid artery of nude mice and harvest of brain metastases, resulting in variants termed MDA-231 BR1, -BR2 and -BR3. The metastasis-selected cells had increased potential for experimental brain metastasis and mice injected with these cells had significantly shorter mean survival than mice injected with the original cell line. Brain metastatic lesions of the selected variants contained significantly more CD31-positive blood vessels than metastases of the non-selected cell line. The variants selected from brain metastases released significantly more VEGF-A and IL-8 into culture supernatants than the original cell line, and more VEGF-A RNA when cultured in normoxic conditions. Mice injected with MDA-231 BR3 into the carotid artery were treated with the VEGF-receptor tyrosine kinase inhibitor PTK787/Z 222584. Oral administration of the inhibitor resulted in a significant decrease in brain tumor burden, reduced CD31-positive vessels in the brain lesions and incidence of PCNA positive tumor cells, and increased apoptosis in the tumor, as measured by TUNEL labeling. We conclude that elevated VEGF expression contributes to the ability of breast cancer cells to form brain metastases. Targeting endothelial cells with a VEGF-receptor specific tyrosine kinase inhibitor reduced angiogenesis and restricted the growth of the brain metastases.

brain metastasis breast cancer tyrosine kinase inhibitor VEGF-A 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Hagemeister FB, Buzdar AV, Luna MA, Blumenschein M. Causes of death in breast cancer: A clinicopathological study. Cancer 1980; 46: 162–7.PubMedCrossRefGoogle Scholar
  2. 2.
    Tsukada Y, Fouad A, Pickren JW, Lane WW. Central nervous system metastasis from breast carcinoma. Autopsy study. Cancer 1983; 52: 2349–54.PubMedCrossRefGoogle Scholar
  3. 3.
    Boogerd W, Vos VW, Hart AAM, Baris G. Brain metastases in breast cancer; natural history, prognostic factors and outcome. J Neuro-Oncol 1993; 15: 165–74.CrossRefGoogle Scholar
  4. 4.
    Freilich RJ, Seidman AD, DeAngelis LM. Central nervous system progression of metastatic breast cancer in patients treated with paclitaxel. Cancer 1995; 76: 232–6.PubMedCrossRefGoogle Scholar
  5. 5.
    Zhang RD, Price JE, Fujimaki T et al. Differential permeability of the blood-brain barrier in experimental brain metastases produced by human neoplasms implanted into nude mice. Am J Pathol 1992; 141: 1115–24.PubMedGoogle Scholar
  6. 6.
    Tikhtman AJ, Patchell RA. Morantz RA, Walsh JW eds. Brain Tumors: A Comprehensive Text. New York: Marcel Dekker 1994; 553–66.Google Scholar
  7. 7.
    Hwang TL, Close TP, Grego JM et al. Predilection of brain metastasis in gray and white matter junction and vascular border zones. Cancer 1996; 77: 1551–5.PubMedCrossRefGoogle Scholar
  8. 8.
    Senger DR, Van De Water L, Brown LFB et al. Vascular permeability factor (VPF,VEGF) in tumor biology. Cancer Metast Rev. 1993; 12:303–24.CrossRefGoogle Scholar
  9. 9.
    Toi H, Hoshina S, Takayanagi T, Tominaga T. Association of vascular endothelial growth factor expression with tumor angiogenesis and with early relapse in primary breast cancer. Jpn J Cancer Res 1994; 85: 1045–9.PubMedGoogle Scholar
  10. 10.
    Samoto K, Ikezaki K, Ono M et al. Expression of vascular endothelial growth factor and its possible relation with neovascularization in human brain tumors. Cancer Res 1995; 55: 1189–93.PubMedGoogle Scholar
  11. 11.
    Strugar J, Rothbart D, Harrington W, Criscuolo GR. Vascular permeability factor in brain metastases: correlation with vasogenic brain edema and tumor angiogenesis. J Neurosurg 1994; 81: 560–6.PubMedCrossRefGoogle Scholar
  12. 12.
    Takano S, Yoshii Y, Kondo S et al. Concentration of vascular endothelial growth factor in the serum and tumor tissue of brain tumor patients. Cancer Res 1996; 56: 2185–90.PubMedGoogle Scholar
  13. 13.
    Mori A, Arii S, Furutani M et al. Vascular endothelial growth factorinduced tumor angiogenesis and tumorigenicity in relation to metastasis in a HT1080 human fibrosarcoma cell model. Int J Cancer 1999; 80: 738–43.PubMedCrossRefGoogle Scholar
  14. 14.
    Potgens AJ, van Altena MC, Lubsen NH et al. Analysis of the tumor vasculature and metastatic behavior of xenografts of human melanoma cell lines transfected with vascular permeability factor. Am J Pathol 1996; 148: 1203–17.PubMedGoogle Scholar
  15. 15.
    Ruiter DJ, Leenders WPJ, Wesseling P et al. Vascular endothelial growth factor-A165 induces progression of melanoma brain metastases without induction of sprouting angiogenesis. Cancer Res 2002; 62: 341–5.PubMedGoogle Scholar
  16. 16.
    Schackert G, Price JE, Bucana CD, Fidler IJ. Unique patterns of brain metastasis produced by different human carcinomas in athymic nude mice. Int J Cancer 1989; 44: 892–7.PubMedGoogle Scholar
  17. 17.
    Zünd G, Uezono S, Stahl GL et al. Hypoxia enhanced induction of endothelial ICAM-1: Role for metabolic acidosis and proteasomes. Am J Physiol 1997; 273: C1571–80PubMedGoogle Scholar
  18. 18.
    Huang S, Robinson JB, DeGuzman A et al. Blockade of NF-κB signalling inhibits angiogenesis and tumorigenicity of human ovarian cancer cells by suppressing expression of vascular endothelial growth factor and interleukin-8. Cancer Res 2000; 60: 5334–9.PubMedGoogle Scholar
  19. 19.
    Huang S, Jean D, LucaMet al. Loss of AP-2 results in downregulation of c-kit and enhancement of melanoma tumorigenicity and metastasis. EMBO J 1998; 17: 4358–69.PubMedCrossRefGoogle Scholar
  20. 20.
    Wood JM, Bold G, Buchdunger E et al. PTK787/ZK222584, a novel and potent inhibitor of vascular endothelial growth factor receptor tyrosine kinases, impairs vascular endothelial growth factor-induced responses and tumor growth after oral administration. Cancer Res 2000; 60: 2178–89.PubMedGoogle Scholar
  21. 21.
    Radinsky R, Fidler IJ, Price JE et al. Terminal differentiation and apoptosis in experimental lung metastases of human osteogenic sarcoma cells by wild type p53. Oncogene 1994; 9: 1877–83.PubMedGoogle Scholar
  22. 22.
    Weidner N, Semple JP, Welch WR, Folkman J. Tumor angiogenesis and metastasis-Correlation in invasive breast carcinoma. N Engl J Med 1991; 324: 1–8.PubMedCrossRefGoogle Scholar
  23. 23.
    Levy AP, Levy NS, Wegner S, Goldberg MA. Transcriptional regulation of the rat vascular endothelial growht factor gene by hypoxia. J Biol Chem 1995; 270: 13333–40.PubMedCrossRefGoogle Scholar
  24. 24.
    Price JE. Metastasis from human breast cancer cell lines. Breast Cancer Res Treat 1996; 39: 93–102.PubMedCrossRefGoogle Scholar
  25. 25.
    Cormack BP, Valdivia R, Falkow S. FACS-optimized mutants of the green fluorescent protein (GFP). Gene 1996; 173: 33–8.PubMedCrossRefGoogle Scholar
  26. 26.
    Chishima T, Miyagi Y, Wang X et al. Cancer invasion and micrometastasis visualized in live tissue by green fluorescent protein expression. Cancer Res 1997; 57: 2042–7.PubMedGoogle Scholar
  27. 27.
    MacDonald TJ, Tabrizi P, Shimada H et al. Detection of brain tumor invasion and micrometastasis in vivo by expression of enhanced green fluorescent protein. Neurosurgery 1998; 43: 1437–42.PubMedCrossRefGoogle Scholar
  28. 28.
    Yang M, Baranov E, Jiang P et al. Whole-body optical imaging of green fluorescent protein-expressing tumors and metastases. Proc Natl Acad Sci USA 2000; 97: 1206–11.PubMedCrossRefGoogle Scholar
  29. 29.
    Migliaccio AR, Bengra C, Ling J et al. Stable and instable transgene integration sites in the human genome: extinction of the Green Fluorescent Protein transgene in K562 cells. Gene 2000; 256: 197–214.PubMedCrossRefGoogle Scholar
  30. 30.
    Liu HS, Jan MS, Chou CK et al. Is green fluorescent protein toxic to the living cells? Biochem Biophys Res Commun 1999; 260: 712–7.PubMedCrossRefGoogle Scholar
  31. 31.
    Yoneda T, Williams PJ, Hiraga T et al. A bone-seeking clone exhibits different biological properties from the MDA-MB-231 parental human breast cancer cells and a brain-seeking clone in vivo and in vitro. J Bone Miner Res 2001; 16: 1486–95.PubMedCrossRefGoogle Scholar
  32. 32.
    Yano S, Shinohara H, Herbst RS et al. Expression of vascular endothelial growth factor is necessary but not sufficient for production and growth of brain metastasis. Cancer Res 2000; 60: 4959–67.PubMedGoogle Scholar
  33. 33.
    Fidler IJ, Yano S, Zhang RD et al. The seed and soil hypothesis: Vascularisation and brain metastases. Lancet Oncol 2002; 3: 53–7.PubMedCrossRefGoogle Scholar
  34. 34.
    De Jong JS, van Diest PJ, van der Valk P, Baak JPA. Expression of growth factors, growth inhibiting factors, and their receptors in invasive breast cancer.I An inventory in search of autocrine and paracrine loops. J Pathol 1998; 184: 44–52.PubMedCrossRefGoogle Scholar
  35. 35.
    De Larco JE, Wuertz BRK, Rosner KA et al. A potential role for interleukin-8 in the metastatic phenotype of breast carcinoma cells. Am J Pathol 2001; 158: 639–46.PubMedGoogle Scholar
  36. 36.
    Gasparini G. Clinical significance of determination of surrogate markers of angiogenesis in breast cancer. Crit Rev Oncol/Hematol 2001; 37: 97–114.CrossRefGoogle Scholar
  37. 37.
    Nör JE, Christensen J, Liu J et al. Up-regulation of Bcl-2 in microvascular endothelial cells enhances intratumoral angiogenesis and accelerates tumor growth. Cancer Res 2001; 61: 2183–8.PubMedGoogle Scholar
  38. 38.
    Bachelder RE, Crago A, Chung J et al. Vascular endothelial growth factor is an autocrine survival factor for neuropilin-expressing breast carcinoma cells. Cancer Res 2001; 61: 5736–40.PubMedGoogle Scholar
  39. 39.
    Soker S, Takashima S, Miao HQ et al. Neuropilin-1 is expressed by endothelial and tumor cells as an isoform-specific receptor for vascular endothelial growth factor. Cell 1998; 92: 735–45.PubMedCrossRefGoogle Scholar
  40. 40.
    Maxwell PH, Dachs GU, Gleadle JM et al. Hypoxia-inducible factor-1 modulates gene expression in solid tumors and influences both angiogenesis and tumor growth. Proc Natl Acad Sci USA 1997; 94: 8104–9.PubMedCrossRefGoogle Scholar
  41. 41.
    Carmeliet P, Dor Y, Herbert JM et al. Role of HIF1-α in hypoxia-mediated apoptosis, cell-proliferation and tumor angiogenesis. Nature 1998; 394: 485–90.PubMedCrossRefGoogle Scholar
  42. 42.
    Knowles HJ, Harris AL. Hypoxia and oxidative stress in breast cancer. Hypoxia and tumourigenesis. Breast Cancer Res 2001; 3: 318–22.PubMedCrossRefGoogle Scholar
  43. 43.
    Xu L, Xie K, Mukaida N et al. Hypoxia-induced elevation in interleukin-8 expression by human ovarian carcinoma cells. Cancer Res 1999; 59: 5822–29.PubMedGoogle Scholar
  44. 44.
    Blancher C, Moore JW, Talks KL et al. Relationship of hypoxia-inducible factor (HIF)-1α and HIF-2α expression to vascular endothelial growth factor induction and hypoxia survival in human breast cancer cell lines. Cancer Res 2000; 60: 7106–13.PubMedGoogle Scholar
  45. 45.
    Bos R, Zhong H, Hanrahan CF et al. Levels of hypoxia-inducible factor-1α during breast carcinogenesis. J Natl Cancer Inst 2001; 93 (4): 309–14.PubMedCrossRefGoogle Scholar
  46. 46.
    Maity A, Pore N, Lee J et al. Epidermal growth factor receptor transcriptionally up-regulates vascular endothelial growth factor expression in human glioblastoma cells via a pathway involving phosphatidylinositol 3′-kinase and distinct from that induced by hypoxia. Cancer Res 2000; 60 (20): 5879–86.PubMedGoogle Scholar
  47. 47.
    Bancroft CC, Chen Z, Dong G et al. Coexpression of proangiogenic factors IL-8 and VEGF by human head and neck squamous cell carcinoma involves coactivation byMEK-MAPK and IKK-NF-κB signal pathways. Clin Cancer Res 2001; 7: 435–42.PubMedGoogle Scholar
  48. 48.
    Yen L, You XL, Al Moustafa AE et al. Heregulin selectively upregulates vascular endothelial growth factor secretion in cancer cells and stimulates angiogenesis. Oncogene 2000; 19 (31): 3460–9.PubMedCrossRefGoogle Scholar
  49. 49.
    O-charoenrat P, Rhys-Evans P, Modjtahedi H, Eccles SA. Vascular endothelial growth factor family members are differentially regulated by c-erbB signaling in head and neck squamous carcinoma cells. Clin Exp Metast 2000; 18 (2): 155–61.CrossRefGoogle Scholar
  50. 50.
    Kanayama H, Yano S, Kim SJ et al. Expression of vascular endothelial growth factor by human renal cancer cells enhances angiogenesis of primary tumors and production of ascites but not metastasis to the lungs of nude mice. Clin Exp Metast 1999; 17: 831–40.CrossRefGoogle Scholar
  51. 51.
    Kim KJ, Li B, Winer J et al. Inhibition of vascular endothelial growth factor-induced angiogenesis suppresses tumor growth in vivo. Nature 1993; 362: 841–4.PubMedCrossRefGoogle Scholar
  52. 52.
    Presta LG, Chen H, O'Connor SJ et al. Humanization of an antivascular endothelial growth factor monoclonal antibody for the therapy of solid tumors and other disorders. Cancer Res 1997; 57: 4593–9.PubMedGoogle Scholar
  53. 53.
    Witte L, Hicklin DJ, Zhu Z et al. Monoclonal antibodies targeting the VEGF receptor-2 (Flk1/KDR) as an anti-angiogenic therapeutic strategy. Cancer Metast Rev 1998; 17: 155–61.CrossRefGoogle Scholar
  54. 54.
    Skobe M, Rockwell P, Goldstein N et al. Halting angiogenesis suppresses carcinoma cell invasion. Nat Med 1997; 3: 1222–7.PubMedCrossRefGoogle Scholar
  55. 55.
    Lin P, Sankar S, Shan S et al. Inhibition of tumor growth by targeting tumor endothelium using a soluble vascular endothelial growth factor receptor. Cell Growth Differ 1998; 9: 49–58.PubMedGoogle Scholar
  56. 56.
    Goldman CK, Kendall RL, Cabrera G et al. Paracrine expression of a native soluble vascular endothelial growth factor receptor inhibits tumor growth,metastasis, and mortality rate. Proc Natl Acad Sci USA 1998; 95: 8795–800.PubMedCrossRefGoogle Scholar
  57. 57.
    Millauer B, Longhi MP, Plate KH et al. Dominant-negative inhibition of Flk-1 suppresses the growth of many tumor types in vivo. Cancer Res 1996; 56: 1615–20.PubMedGoogle Scholar
  58. 58.
    Shibuya M. Role of VEGF-flt receptor system in normal and tumor angiogenesis. Adv Cancer Res 1995; 67: 281–316.PubMedCrossRefGoogle Scholar
  59. 59.
    Yano S, Herbst RS, Shinohara H et al. Treatment for malignant pleural effusion of human lung adenocarcinoma by inhibition of vascular endothelial growth factor receptor tyrosine kinase phosphorylation. Clin Cancer Res 2000; 6: 957–65.PubMedGoogle Scholar
  60. 60.
    Solorzano CC, Baker CH, Bruns CJ et al. Inhibition of growth and metastasis of human pancreatic cance growing in nude mice by PTK 787/ZK222584, and inhibitor of the vascular endothelial growth factor receptor tyrosine kinases. Cancer Biother Radiopharm 2001; 16: 359 70.PubMedCrossRefGoogle Scholar
  61. 61.
    Xu L, Yoneda J, Herrera C et al. Inhibition of malignant ascites and growth of human ovarian carcinoma by oral administration of a potent inhibitor of the vascular endothelial growth factor receptor tyrosine kinases. Int J Oncol 2000; 16: 445–54.PubMedGoogle Scholar
  62. 62.
    Lin B, Podar K, Gupta D et al. The vascular endothelial growth factor receptor tyrosine kinase inhibitor PTK787/ZK222584 inhibits growth and migration of multiple myeloma cells in the bone marrow microenvironment. Cancer Res 2002; 62: 5019–26.PubMedGoogle Scholar

Copyright information

© Kluwer Academic Publishers 2004

Authors and Affiliations

  • Lee Su Kim
    • 1
  • Suyung Huang
    • 2
  • Weixin Lu
    • 2
  • Dina Chelouche Lev
    • 2
  • Janet E. Price
    • 2
  1. 1.Department of Surgery, Han-gang Sacred Heart Hospital, College of MedicineHallym UniversitySeoulSouth Korea
  2. 2.Department of Cancer BiologyUniversity of Texas M.D. Anderson Cancer CenterHoustonUSA

Personalised recommendations