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Surrogate Markers of Angiogenesis and Metastasis

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Metastasis Research Protocols

Part of the book series: Methods in Molecular Medicine ((MIMM,volume 57))

Abstract

Angiogenesis, the formation of new blood vessels from a preexisting vascular network, is a complex multistep process under the control of positive and negative factors (bi1,bi2). Growth, progression, and metastasis of malignant tumors are angiogenesis-dependent processes (3-bi5). There is compelling evidence that development of angiogenic phenotype is a common pathway in the biology of tumor progression and metastasis, and the angiogenesis is correlated with other molecular mechanisms (e.g., the control of wild-type p53 on vascular endothelial growth factor or thrombospondin-1) (6,7). Angiogenesis can be directly determined, and so it may be useful as a biomarker (8-bi10). Tumor microvessel density (11-13) or tissue angiogenic factors expression may predict which patients are at different risk of metastasis (14-16), and serum levels of angiogenic factors may serve as tumor markers (17,18).

At present the most widely used method to assess angiogenesis is quantification of intratumoral microvessel density (IMD) of the primary tumor by using specific markers for endothelial cells such as factor VIII-related antigen (FVIII-RA), anti-CD31 (platelet endothelial cell adhesion molecule, PECAM), or anti-CD34 antibodies (19,22), using a standard immunocytochemistry immunoperoxidase technique to stain micro vessels (23,24). The basic principles of immunocytochemistry are described in Chapter 2 by Brooks and use of immunocytochemistry employing antibodies against factor VIII-related antigen and CD31 is described in Chapter 8 by Turner and Harris.

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References

  1. Folkman, J. (1992) The role of angiogenesis in tumour growth. Semin. Cancer Biol. 3, 65–72.

    CAS  PubMed  Google Scholar 

  2. Folkman, J. (1993) Tumor angiogenesis, in Cancer Medicine,4th ed. (Holland, J., Frei, E., and Bast, R. C., eds.), Lea and Febiger, Melbourne, 153–171.

    Google Scholar 

  3. Folkman, J. (1990) What is the evidence that tumors are angiogenesis dependent? J. Natl. Cancer Inst. 82,4–6.

    Article  CAS  PubMed  Google Scholar 

  4. D'Amore, P. A. (1992) Capillary growth: a two-cell system. Cancer Biol. 3,49–56.

    Google Scholar 

  5. D'Amore, P. A. and Smith, S. R. (1993) Growth factors effects on cells of the vascular wall: a survey. Growth Factors 8,61–75.

    Article  Google Scholar 

  6. Dameron, K. M., Volpert, O. V., Tainsky, M. A., and Bouck, N. (1994) Control of angiogenesis in fibroblasts by p53 regulation of thrombospondin-1. Science 265, 1582–1585.

    Article  CAS  PubMed  Google Scholar 

  7. Kieser, A., Wiech, H. A., Bradner, G., Marmé, D., and Kolch, W. (1994) Mutant p53 potentiates protein kinase C induction of vascular endothelial growth factor expression. Oncogene 9,63–969.

    Google Scholar 

  8. Gasparini, G. (1996) Clinical significance of the determination of angiogenesis inhuman breast cancer: update of the biological background and overview of the Vicenza studies. Eur. J. Cancer 32A, 2485–2493.

    Article  CAS  PubMed  Google Scholar 

  9. Gasparini, G. and Harris, A. L. (1999) Prognostic significance of tumour vascu-larity, in Antiangiogenic Agents in Cancer Therapy (Teicher, B. A.,ed.), Humana Press, Totowa, NJ, 317–319.

    Google Scholar 

  10. Gasparini, G. and Harris, A. L. (1995) Clinical importance of the determination of tumor angiogenesis in breast carcinoma: much more than a new prognostic tool. J. Clin. Oncol. 13, 765–782.

    CAS  PubMed  Google Scholar 

  11. Gasparini, G. (1994) Quantification of intratumoral vascularization predicts metastasis in human invasive solid tumors. Oncol. Rep. 1, 7–12.

    Google Scholar 

  12. Gasparini, G. (1997) Prognostic and predictive value of intra-tumoral microvessel density in human solid tumors, in Tumor Angiogenesis (Bicknell, R., Lewis, C. E., Ferrara, M., eds.), Oxford University Press, Oxford, 29–44.

    Google Scholar 

  13. De Jong, J. S., Van Diest, P. J., and Baak, J. P.A. (1995) Methods in laboratory investigation. Heterogeneity and reproducibility of microvessel counts in breast cancer. Lab. Invest. 73, 922–926.

    PubMed  Google Scholar 

  14. Ambs, S., Bennett, W. P., Merriam, W. G., Ogunfusika, M. O., Oser, S. M., Khan, M. A., and Harris, C. C. (1998) Vascular endothelial growth factor and nitric oxide synthase expression in human lung cancer and the relation to p53. Br. J. Cancer 78, 233–239.

    Article  CAS  PubMed  Google Scholar 

  15. Gasparini, G., Fox, S. B., Verderio, P., Bonoldi, E., Boracchi, P., et al. (1996) Determination of angiogenesis adds information to estrogen receptor status in predicting the effacy of adjuvant tamoxifen in node-positive breast cancer patients. Clin. Cancer Res. 2,1191–1198.

    CAS  PubMed  Google Scholar 

  16. Gasparini, G., Bonoldi, E., Viale, G., Verderio, P., Boracchi, P., Panizzoni, G. A., et al. (1996) Prognostic and predictive value of tumor angiogenesis in ovarian carcinomas. Int. J. Cancer (Pred. Oncol.) 69,205–211.

    Article  CAS  Google Scholar 

  17. Vermeulen, P. B., Gasparini, G., Fox, S. B., Toi, M., Martin, L., McCulloch, P., et al. (1996) Quantification of angiogenesis in solid human tumours: an international consensus on the methodology and criteria of evaluation. Eur. J. Cancer 32,2478–2484.

    Article  Google Scholar 

  18. Obermair, A., Tempfer, C., Hefler, L., Preyer, O., Kaider, A., Zeillinger, R., et al. (1998) Concentration of vascular endothelial growth factor (VEGF) in the serum of patients with suspected ovarian cancer. Br. J. Cancer 77, 1870–1874.

    Article  CAS  PubMed  Google Scholar 

  19. Parums, D. V., Cordell, J. L., Micklem, K., Heryet, A. R., Gatte, K. C., and Mason, D. Y. (1990) JC70: a new monoclonal antibody that detects vascular endothelium associated antigen on routinely processed tissue sections. J. Clin. Pathol. 43,752–757.

    Article  CAS  PubMed  Google Scholar 

  20. Horak, E., Leek, R., Klenk, N., Lejeune, S., Smith, K., Stuart, N., et al. (1992) Angiogenesis, assessed by platelet/endothelial cell adhesion molecule antibodies, as an indicator of node metastasis and survival in breast cancer. Lancet 340,1120–1124.

    Article  CAS  PubMed  Google Scholar 

  21. Ruiter, D. J., Schlingemann, R. O., Rietveid, F. J. R., and de Waal, R. M. W. (1989) Monoclonal antibody-defined human endothelial antigens as a vascular marker. Invest Dermatol. 93, 25–32.

    Article  Google Scholar 

  22. Traweek, S. T., Kandalaft, P. L., Mehta, P., and Battifora, H. (1991) The human hematopoietic progenitor cell antigen (CD34) in vascular neoplasia. Am. J. Clin. Pathol. 96, 25–31.

    CAS  PubMed  Google Scholar 

  23. Sternberger, S. S. and De Lellis, R. A., eds. (1982) Diagnostic Immunohisto-chemistry, Masson, New York.

    Google Scholar 

  24. Bullock, G. R. and Petrusz, P., eds. (1982 and 1983) Techniques in Immunochem-istry, vols. 1 and 2. Academic, London.

    Google Scholar 

  25. Gasparini, G., Brooks, P. C., Biganzoli, E., Vermeulen, P. B., and Dirix, L. Y., and Ranieri, G., et al. (1998) Vascular integrin avβ3: a new prognostic indicator in breast cancer. Clin. Cancer Res. 4,1264–2625.

    Google Scholar 

  26. Weidner, N., Semple, J. P., Welch, W. R., and Folkman, J. (1991) Tumor angiogenesis and metastasis-correlation in invasive breast carcinoma. N. Engl. J. Med. 324,1–8.

    Article  CAS  PubMed  Google Scholar 

  27. Fox, S. B., Leek, R. D., Weekes, M. P., Withehouse, R. M., Gatter, K. C., and Harris, A. L. (1995) Quantitation and prognostic value of breast cancer angiogenesis: comparison of microvessel density, Chalkey count and computer image analysis. J. Pathol. 77,277–283.

    Google Scholar 

  28. Visscher, D., Smilanetz, S., Drozdowicz, S., Wilkes, S. (1993) Prognostic significance of image morphometric microvessel enumeration in breast carcinoma. Anal. Quant. Cytol. 15, 88–92.

    CAS  Google Scholar 

  29. Simpson, J. F., Ahn, C., Battifora, M., and Esteban, J. M. (1996) Endothelial area as a prognostic indicator for invasive breast carcinoma. Cancer 77,2077–2085.

    Article  CAS  PubMed  Google Scholar 

  30. Barbareschi, M., Weidner, N., Gasparini, G., Morelli, L., Forti, S., Eccher, C., et al. (1995) Microvessel density quantification in breast carcinomas. Assessment by light microscopy vs computers aided image analysis system. Appl. Immunohistochem. 3, 75–84.

    Google Scholar 

  31. Barth, P. J., Weingartner, K., Kohler, H. H., and Bittinger, A. (1996) Assessment of the vascularization in prostatic carcinoma: a morphometric investigation. Hum. Pathol. 27,1306–1310.

    Article  CAS  PubMed  Google Scholar 

  32. Toi, M., Ueno, T., Matsumoto, H., Saji, H., Funata, N., Koike, M., and Tominaga, T. (1999) Significance of thymidine phosphorylase as a marker of tumor mono-cytes in breast cancer. Clin. Cancer Res. 5, 1131–1137.

    CAS  PubMed  Google Scholar 

  33. Toi, M., Hoshina, S., Taniguchi, T., Yamamoto, Y., Ishitsuka, H., and Tominaga, T. (1995) Expression of platelet-derived endothelial cell growth factor/thymidine phoshorylase in human breast cancer. Int. J. Cancer (Pred. Oncol.) 64,79–82.

    Article  CAS  Google Scholar 

  34. Fox, S. B., Engels, K., Comley, M., Witehouse, R. M., Turley, H., Gatter, K. C., and Harris, A. L. (1997) Relationship of elevated tumour thymidine phosphorylase in node-positive breast carcinomas to the effects of adjuvant CMF. Ann. Oncol. 8,271–275.

    Article  CAS  PubMed  Google Scholar 

  35. Fox, S. B., Westwood, M., Moghaddam, A., Comley, M., Turley, H., Witehouse, R. M., et al. (1996) The angiogenic factor platelet-derived endothelial cell growth factor/hymidine phosphorylase is up-regulated in breast cancer epithelium and endothelium. Br. J. Cancer 73, 275–280.

    Article  CAS  PubMed  Google Scholar 

  36. Polak, J. M. and van Noorden, S., eds. (1983) Immunocytochemistry. John Wright and Sons, Bristol, UK.

    Google Scholar 

  37. Nagy, J., Curry, G. W., Hillan, K. J., Purushotham, A. D., and George, W. D. (1996) Hepatocyte growth factor/scatter factor, angiogenesis and tumour cell proliferation in primary breast cancer. Breast 5, 105–109.

    Article  Google Scholar 

  38. Colomer, R., Aparicio, J., Montero, S., Guzmán, C., Larrodera, L., and Cortes-Funes, H. (1997) Low levels of basic fibroblast growth factor (bFGF) are associated with a poor prognosis in human breast carcinoma. Br. J. Cancer 76,1215–1220.

    Article  CAS  PubMed  Google Scholar 

  39. Gasparini, G., Toi, M., Miceli, R., Vermeulen, P. B., Dittadi, R., Biganzoli, E., et al. (1999) Clinical relevance of vascular endothelial growth factor and thymidine phosphorylase in patients with node-positive breast cancer treated with either adjuvant chemotherapy or hormone therapy. Cancer J. Sci. Am. 5,101–111.

    CAS  PubMed  Google Scholar 

  40. Hyodo, I., Doi, T., Endo, H., Hosokawa, Y., Nishikawa, Y., Tanimizu, M., et al. (1998) Clinical significance of plasma vascular endothelial growth factor in gastrointestinal cancers. Eur. J. Cancer 34, 2041–2045.

    Article  CAS  PubMed  Google Scholar 

  41. Toi, M., Taniguchi, T., Yamamoto, Y., Kurisaki, T., Suzuki, H., and Tominaga, T. (1996) Clinical significance of the determination of angiogenic factors. Eur. J. Cancer 32A, 2513–2519.

    Article  CAS  PubMed  Google Scholar 

  42. Morelli, D., Lazzerini, D., Cazzaniga, S., Squicciarini, P., Bignami, P., Maier, J. A., et al. (1998) Evaluation of the balance between angiogenic and antiangiogenic circulating factors in patients with breast and gastrointestinal cancers. Clin. Cancer Res. 4, 1221–1225.

    CAS  PubMed  Google Scholar 

  43. Salven, P., Orpana, A., and Joensuu, H. (1999) Leukocytes and platelets of patients with cancer contain high levels of vascular endothelial growth factor. Clin. Cancer Res. 5,487–491.

    CAS  PubMed  Google Scholar 

  44. Banks, R. E., Forbes, M. A., Kinsey, S. E., Stanley, A., Ingham, E., Walters, C., and Selby, P. J. (1998) Release of the angiogenic cytokine vascular endothelial growth factor (VEGF) from platelets: significance for VEGF measurements and cancer biology. Br. J. Cancer 77, 956–964.

    Article  CAS  PubMed  Google Scholar 

  45. Wynendaele, W., Derua, R., Hoylaerts, M. F., Pawinski, A., Waelkens, E., Bruijn, E. A., et al. (1999) Vascular endothelial growth factor measured in platelet poor plasma allows optimal separation between cancer patients and volunteers: a key to study an angiogenic marker in vivo? Ann. Oncol. 10, 965–971.

    Article  CAS  PubMed  Google Scholar 

  46. Waterborg, J. H. and Matthews, H. R. (1984) The Lowry method for protein quantitation, in Methods in Molecular Biology, vol. 1 (Walker, J., ed.), Humana Press, Totowa, NJ, 1–3.

    Google Scholar 

  47. Fontanini, G., Vignati, S., Lucchi, M., Mussi, A., Calcinai, A., Boldrini, L., et al. (1997) Neoangiogenesis and p53 protein in lung cancer: their prognostic role and their relation with vascular endothelial growth factor (VEGF) expression. Br. J. Cancer 75, 1295–1301.

    Article  CAS  PubMed  Google Scholar 

  48. DePlacido, S., Carlomagno, C., Ciardiello, F., DeLaurentiis, M., Pepe, S., Ruggiero, A., et al. (1999) Measurement of neovascularization is an independent prognosticator of survival in node-negative breast cancer patients with long-term follow-up. Clin. Cancer Res. 5,2854–2859.

    CAS  Google Scholar 

  49. Dickinson, A. J., Fox, S. B., Persad, R. A., Hollyer, J., Sibley, G. N. A., and Harris, A. L. (1994) Quantitation of angiogenesis as an independent predictor of prognosis in invasive bladder carcinomas. Br. J. Urol. 74, 762–766.

    Article  CAS  PubMed  Google Scholar 

  50. Gasparini, G., Toi, M., Gion, M., Verderio, P., Dittadi, R., Hanatani, M., et al. (1997) Prognostic significance of vascular endothelial growth factor protein in node-negative breast carcinoma. J. Natl. Cancer Inst. 89, 139–147.

    Article  CAS  PubMed  Google Scholar 

  51. Toi, M., Gion, M., Biganzoli, E., Dittadi, R., Boracchi, P., Miceli, R., et al. (1997) Co-determination of the angiogenic factors thymidine phosphorylase and vascular endothelial growth factor in node-negative breast cancer: prognostic implications. Angiogenesis 1, 71–83.

    Article  CAS  PubMed  Google Scholar 

  52. Linderholm, B., Tavelin, B., Grankvist, K., and Henriksson, R. (1998) Vascular endothelial growth factor is of high prognostic value in node-negative breast carcinoma. J. Clin. Oncol. 16, 3121–3128.

    CAS  PubMed  Google Scholar 

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Author information

Authors and Affiliations

  1. Division of Medical Oncology, Azienda Ospedali Riuniti, Reggio Calabria, Italy

    Girolamo Ranieri

  2. Division of Medical Oncology, Azienda Complesso Ospedaliero “S. Filippo Neri,”, Rome, Italy

    Giampietro Gasparini

Authors
  1. Girolamo Ranieri
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  2. Giampietro Gasparini
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Editor information

Editors and Affiliations

  1. Research School of Biological and Molecular Sciences, Oxford Brookes University, Oxford, UK

    Susan A. Brooks

  2. Institute for Anatomy, Department of Neuroanatomy, University Hospital Hamburg-Eppendorf, Hamburg, Germany

    Udo Schumacher

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Ranieri, G., Gasparini, G. (2001). Surrogate Markers of Angiogenesis and Metastasis. In: Brooks, S.A., Schumacher, U. (eds) Metastasis Research Protocols. Methods in Molecular Medicine, vol 57. Humana Press. https://doi.org/10.1385/1-59259-136-1:99

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  • DOI: https://doi.org/10.1385/1-59259-136-1:99

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-610-9

  • Online ISBN: 978-1-59259-136-7

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