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. 2003 Jun;7(2):57–61. doi: 10.1080/13577140310001607275

Immunohistochemistry Evaluation of the Effect in Vivo of Tumor Necrosis Factor (TNF)-α on Blood Vessel Density in Murine Fibrosarcoma

Avi Eisenthal 1,✉, Ignat Schwartz 1, Josephine Issakov 1, Yossef Klausner 2, Faina Misonzhnik 1, Beatriz Lifschitz-Mercer 1
PMCID: PMC2395522  PMID: 18521369

Abstract

Purpose: Angiogenesis is essential for tumor growth and metastases, thus bestowing obvious importance upon methodologies which could enable its inhibition.

Materials: C57BL/6 female mice bearing a subcutaneous (s.c.) MCA205 fibrosarcoma were used.

Methods: Ten mice were divided equally into two groups. One group was injected intraperitoneally (i.p.) with 10 μg tumor necrosis factor-α (TNF-α and the other (controls) with Hanks balanced salt solution (HBSS). Tumor growth was monitored at least twice weekly. The number of endothelial cells in the blood microvessels was assessed by immunohistostaining on paraffin-embedded tumor tissue sections using vascular endothelial growth factor (VEGF) and Factor 8 antibodies. Expression of the p53 gene was similarly assessed by immunohistostaining.

Results: Injection of 10 μg TNF-α into the tumor-bearing mice reduced the number of endothelial cells in the blood microvessels by 46% on day 3 post-injection which was accompanied by an increase (by 37%) in the expression of p53 in these cells. It also inhibited tumor growth compared to the HBSS-injected group starting at 17 days post-cytokine injection.

Discussion: The antitumor in vivo effect exerted by TNF-α on established murine sarcoma s.c. tumors may be due to an earlier effect of the cytokine on the tumor's blood microvessels, probably through an apoptotic mechanism involving the p53 gene.

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Selected References

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  1. Asher A., Mulé J. J., Reichert C. M., Shiloni E., Rosenberg S. A. Studies on the anti-tumor efficacy of systemically administered recombinant tumor necrosis factor against several murine tumors in vivo. J Immunol. 1987 Feb 1;138(3):963–974. [PubMed] [Google Scholar]
  2. Brem H., Folkman J. Analysis of experimental antiangiogenic therapy. J Pediatr Surg. 1993 Mar;28(3):445–451. doi: 10.1016/0022-3468(93)90246-h. [DOI] [PubMed] [Google Scholar]
  3. Carswell E. A., Old L. J., Kassel R. L., Green S., Fiore N., Williamson B. An endotoxin-induced serum factor that causes necrosis of tumors. Proc Natl Acad Sci U S A. 1975 Sep;72(9):3666–3670. doi: 10.1073/pnas.72.9.3666. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. De Assis M. C., Da Costa A. O., Barja-Fidalgo T. C., Plotkowski M. C. Human endothelial cells are activated by interferon-gamma plus tumour necrosis factor-alpha to kill intracellular Pseudomonas aeruginosa. Immunology. 2000 Oct;101(2):271–278. doi: 10.1046/j.1365-2567.2000.00102.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Eisenthal A., Polyvkin N., Bramante-Schreiber L., Misonznik F., Hassner A., Lifschitz-Mercer B. Expression of dendritic cells in ovarian tumors correlates with clinical outcome in patients with ovarian cancer. Hum Pathol. 2001 Aug;32(8):803–807. doi: 10.1053/hupa.2001.26455. [DOI] [PubMed] [Google Scholar]
  6. Feleszko W., Bałkowiec E. Z., Sieberth E., Marczak M., Dabrowska A., Giermasz A., Czajka A., Jakóbisiak M. Lovastatin and tumor necrosis factor-alpha exhibit potentiated antitumor effects against Ha-ras-transformed murine tumor via inhibition of tumor-induced angiogenesis. Int J Cancer. 1999 May 17;81(4):560–567. doi: 10.1002/(sici)1097-0215(19990517)81:4<560::aid-ijc10>3.0.co;2-7. [DOI] [PubMed] [Google Scholar]
  7. Gutman M., Sofer D., Lev-Chelouche D., Merimsky O., Klausner J. M. Synergism of tumor necrosis factor-alpha and melphalan in systemic and regional administration: animal study. Invasion Metastasis. 1997;17(4):169–175. [PubMed] [Google Scholar]
  8. Hanahan D., Folkman J. Patterns and emerging mechanisms of the angiogenic switch during tumorigenesis. Cell. 1996 Aug 9;86(3):353–364. doi: 10.1016/s0092-8674(00)80108-7. [DOI] [PubMed] [Google Scholar]
  9. Joseph I. B., Isaacs J. T. Macrophage role in the anti-prostate cancer response to one class of antiangiogenic agents. J Natl Cancer Inst. 1998 Nov 4;90(21):1648–1653. doi: 10.1093/jnci/90.21.1648. [DOI] [PubMed] [Google Scholar]
  10. Leek R. D., Hunt N. C., Landers R. J., Lewis C. E., Royds J. A., Harris A. L. Macrophage infiltration is associated with VEGF and EGFR expression in breast cancer. J Pathol. 2000 Mar;190(4):430–436. doi: 10.1002/(SICI)1096-9896(200003)190:4<430::AID-PATH538>3.0.CO;2-6. [DOI] [PubMed] [Google Scholar]
  11. Mazanet R., Antman K. H. Sarcomas of soft tissue and bone. Cancer. 1991 Aug 1;68(3):463–473. doi: 10.1002/1097-0142(19910801)68:3<463::aid-cncr2820680304>3.0.co;2-e. [DOI] [PubMed] [Google Scholar]
  12. McIntosh J. K., Mulé J. J., Merino M. J., Rosenberg S. A. Synergistic antitumor effects of immunotherapy with recombinant interleukin-2 and recombinant tumor necrosis factor-alpha. Cancer Res. 1988 Jul 15;48(14):4011–4017. [PubMed] [Google Scholar]
  13. Nathan I., Dizdaroglu M., Bernstein L., Junker U., Lee C., Muegge K., Durum S. K. Induction of oxidative DNA damage in u937 cells by TNF or anti-Fas stimulation. Cytokine. 2000 Jul;12(7):881–887. doi: 10.1006/cyto.1999.0638. [DOI] [PubMed] [Google Scholar]
  14. Ogata N., Yamamoto H., Kugiyama K., Yasue H., Miyamoto E. Involvement of protein kinase C in superoxide anion-induced activation of nuclear factor-kappa B in human endothelial cells. Cardiovasc Res. 2000 Jan 14;45(2):513–521. doi: 10.1016/s0008-6363(99)00364-8. [DOI] [PubMed] [Google Scholar]
  15. Okuyama M., Yamaguchi S., Yamaoka M., Nitobe J., Fujii S., Yoshimura T., Tomoike H. Nitric oxide enhances expression and shedding of tumor necrosis factor receptor I (p55) in endothelial cells. Arterioscler Thromb Vasc Biol. 2000 Jun;20(6):1506–1511. doi: 10.1161/01.atv.20.6.1506. [DOI] [PubMed] [Google Scholar]
  16. Palladino M. A., Jr, Shalaby M. R., Kramer S. M., Ferraiolo B. L., Baughman R. A., Deleo A. B., Crase D., Marafino B., Aggarwal B. B., Figari I. S. Characterization of the antitumor activities of human tumor necrosis factor-alpha and the comparison with other cytokines: induction of tumor-specific immunity. J Immunol. 1987 Jun 1;138(11):4023–4032. [PubMed] [Google Scholar]
  17. Pluda J. M. Tumor-associated angiogenesis: mechanisms, clinical implications, and therapeutic strategies. Semin Oncol. 1997 Apr;24(2):203–218. [PubMed] [Google Scholar]
  18. Soengas M. S., Alarcón R. M., Yoshida H., Giaccia A. J., Hakem R., Mak T. W., Lowe S. W. Apaf-1 and caspase-9 in p53-dependent apoptosis and tumor inhibition. Science. 1999 Apr 2;284(5411):156–159. doi: 10.1126/science.284.5411.156. [DOI] [PubMed] [Google Scholar]
  19. Torisu H., Ono M., Kiryu H., Furue M., Ohmoto Y., Nakayama J., Nishioka Y., Sone S., Kuwano M. Macrophage infiltration correlates with tumor stage and angiogenesis in human malignant melanoma: possible involvement of TNFalpha and IL-1alpha. Int J Cancer. 2000 Jan 15;85(2):182–188. [PubMed] [Google Scholar]
  20. Volpert O. V., Lawler J., Bouck N. P. A human fibrosarcoma inhibits systemic angiogenesis and the growth of experimental metastases via thrombospondin-1. Proc Natl Acad Sci U S A. 1998 May 26;95(11):6343–6348. doi: 10.1073/pnas.95.11.6343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Yoshimura H., Chikamoto A., Honda T., Tashiro K., Nakamoto T., Takano M., Takagi K., Nagasue N., Soma G. Relationship between microvessel quantification and inducibility of endogenous tumor necrosis factor in colorectal adenocarcinoma. Anticancer Res. 2000 Jan-Feb;20(1C):629–633. [PubMed] [Google Scholar]

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