Immunohistochemical Identification And Localization Of Endogenous Endostatin And Its Related Peptides In Murine Tumors

  • Jianzhong Sun
  • Ivan Ding
  • Bruce Fenton
  • Won Sam Yi
  • Paul Okunieff
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 599)


Endostatin, a fragment of the C-terminal domain of mouse collagen XVIII, is a recently demonstrated endogenous inhibitor of tumor angiogenesis. Although endostatin can be detected in blood and urine of tumor-bearing as well as normal mice, the exact localization of the endogenous protein and its related peptides in tumor tissues is unknown. We used immunohistochemistry and immunoblotting to identify endostatin tissue location and staining patterns in tumor, as well as to determine the differences in the levels of endostatin expression between tumor cells (in vitro) and tumor tissues (in vivo). Using a specific polyclonal antibody against murine endostatin, we quantitatively determined the levels of endostatin in five murine mammary tumors and the KHT sarcoma by Western blotting. The staining patterns for this protein in tumor sections were examined histologically by immunohistochemistry. Our results show that: 1) Endogenous endostatin and its related peptides are widely distributed in all in vivo tumor types tested, but not in most of the cultured tumor cell lines. 2) Endogenous endostatin stained most tumor stromal components, including vessel walls, basement membranes, extracellular spaces, and tumor cells. 3) Staining patterns and localization of endostatin and thrombospondin-1 were similar in these tumor sections.


Related Peptide Collagen Xviii Murine Mammary Tumor Cellular Staining Related Fragment 
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  1. 1.
    J. C. Bout, F. Soncin, B. Vandenbunder, Therapeutic prospects of angiogenesis inhibitors in cancerology, Rev. Mal. Respir. 16, 854-855 (1999).PubMedGoogle Scholar
  2. 2.
    E. Keshet, S. A. Ben-Sasson, Anticancer drug targets: approaching angiogenesis, J. Clin. Invest. 104, 1497-1501 (1999).PubMedCrossRefGoogle Scholar
  3. 3.
    Y. Cao, Therapeutic potentials of angiostatin in the treatment of cancer, Haematologica 84, 643-650 (1999).PubMedGoogle Scholar
  4. 4.
    J. Folkman, Angiogenesis in cancer, vascular, rheumatoid and other disease, Nat. Med. 1, 27-31 (1995).PubMedCrossRefGoogle Scholar
  5. 5.
    J. Folkman, Angiogenesis inhibitors generated by tumors, Mol. Med. 1, 120-122 (1995).PubMedGoogle Scholar
  6. 6.
    P. Hahnfeldt, D. Panigrahy, J. Folkman, L. Hlatky, Tumor development under angiogenic signaling: a dynamical theory of tumor growth, treatment response, and postvascular dormancy, Cancer Res. 59, 4770-4775 (1999).PubMedGoogle Scholar
  7. 7.
    A. Le Querrec, D. Duval, G. Tobelem, Tumour angiogenesis, Baillieres Clin. Haematol. 6, 711-730 (1993).PubMedCrossRefGoogle Scholar
  8. 8.
    M. S. O’Reilly, T. Boehm, Y. Shing, N. Fukai, G. Vasios, W. S. Lane, E. Flynn, J. R. Birkhead, B. R. Olsen, J. Folkman, Endostatin: an endogenous inhibitor of angiogenesis and tumor growth, Cell 88, 277-285 (1997).PubMedCrossRefGoogle Scholar
  9. 9.
    T. Sasaki, H. Larsson, J. Kreuger, M. Salmivirta, L. Claesson-Welsh, U. Lindahl, E. Hohenester, R. Timpl, Structural basis and potential role of heparin/heparan sulfate binding to the angiogenesis inhibitor endostatin, Embo. J. 18, 6240-6248 (1999).PubMedCrossRefGoogle Scholar
  10. 10.
    W. Wen, M. A. Moses, D. Wiederschain, J. L. Arbiser, J. Folkman, The generation of endostatin is mediated by elastase, Cancer Res. 59, 6052-6056 (1999).PubMedGoogle Scholar
  11. 11.
    T. Sasaki, N. Fukai, K. Mann, W. Gohring, B. R. Olsen, R. Timpl, Structure, function and tissue forms of the C-terminal globular domain of collagen XVIII containing the angiogenesis inhibitor endostatin, Embo. J. 17, 4249-4256 (1998).PubMedCrossRefGoogle Scholar
  12. 12.
    M. Dhanabal, R. Ramchandran, M. J. Waterman, H. Lu, B. Knebelmann, M. Segal, V. P. Sukhatme, Endostatin induces endothelial cell apoptosis, J. Biol. Chem. 274, 11721-11726 (1999).PubMedCrossRefGoogle Scholar
  13. 13.
    M. Dhanabal, R. Ramchandran, R. Volk, I. E. Stillman, M. Lombardo, M. L. Iruela-Arispe, M. Simons, V. P. Sukhatme, Endostatin: yeast production, mutants, and antitumor effect in renal cell carcinoma, Cancer Res. 59 , 189-197 (1999).PubMedGoogle Scholar
  14. 14.
    L. Taddei, P. Chiarugi, L. Brogelli, P. Cirri, L. Magnelli, G. Raugei, M. Ziche, H. J. Granger, V. Chiarugi, G. Ramponi, Inhibitory effect of full-length human endostatin on in vitro angiogenesis, Biochem. Biophys. Res. Commun. 263, 340-345 (1999).Google Scholar
  15. 15.
    P. Blezinger, J. Wang, M. Gondo, A. Quezada, D. Mehrens, M. French, A. Singhal, S. Sullivan, A. Rolland, R. Ralston, W. Min, Systemic inhibition of tumor growth and tumor metastases by intramuscular administration of the endostatin gene, Nat. Biotechnol. 17, 343-348 (1999).PubMedCrossRefGoogle Scholar
  16. 16.
    T. Boehm, J. Folkman, T. Browder, M. S. O’Reilly, Antiangiogenic therapy of experimental cancer does not induce acquired drug resistance see comments, Nature 390, 404-407 (1997).PubMedCrossRefGoogle Scholar
  17. 17.
    Z. Chang, A. Choon, A. Friedl, Endostatin binds to blood vessels in situ independent of heparan sulfate and does not compete for fibroblast growth factor-2 binding, Am. J. Pathol. 155, 71-76 (1999).PubMedGoogle Scholar
  18. 18.
    D. Schuppan, T. Cramer, M. Bauer, T. Strefeld, E. G. Hahn, H. Herbst, Hepatocytes as a source of collagen type XVIII endostatin letter, Lancet 352, 879-880 (1998).PubMedCrossRefGoogle Scholar
  19. 19.
    H. Suit, A. Allam, J. Allalunis-Turner, W. Brock, T. Girinsky, S. Hill, N. Hunter, L. Milas, R. Pearcey, L. Peters, et al. Is tumor cell radiation resistance correlated with metastatic ability? Cancer Res. 54(7):1736-1741 (1994).PubMedGoogle Scholar

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© Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • Jianzhong Sun
    • 1
  • Ivan Ding
    • 1
  • Bruce Fenton
    • 1
  • Won Sam Yi
    • 1
  • Paul Okunieff
    • 1
  1. 1.Department of Radiation OncologyUniversity of Rochester School of MedicineRochesterUSA

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