Histochemistry and Cell Biology

, Volume 122, Issue 1, pp 27–33 | Cite as

Anti-angiogenic effects of thalidomide: expression of apoptosis-inducible active-caspase-3 in a three-dimensional collagen gel culture of aorta

  • Keiko Fujita
  • Yoshiko Asami
  • Kayoko Tanaka
  • Masumi Akita
  • Hans-Joachim Merker
Original Paper
First Online:
Accepted:

Abstract

The anti-angiogenic properties of thalidomide have led to the use of the agent as a remedy for multiple myeloma. Nevertheless, the anti-angiogenic moiety of thalidomide remains unidentified. In this study we examined the anti-angiogenic effects of thalidomide in an in vitro model using a three-dimensional collagen gel culture. Angiogenesis was significantly inhibited when the culture was treated with thalidomide plus cytochrome P-450 (CYP2B4), and the migrating cells and tubules were positive for active-caspase-3 in an accompanying immunohistochemical investigation. Transmission electron microscopic observation also confirmed that active-caspase-3-positive cells demonstrated apoptotic characteristics. This study is the first to morphologically demonstrate the effect of thalidomide in directly inducing the apoptosis of new tubules and migrating cells on a three-dimensional collagen gel culture of aorta. Taken together with earlier findings, our new results indicate that the thalidomide-induced inhibition of angiogenesis involves apoptosis in addition to the suppression of TNF-α and inhibition of cell migration from aorta explants, i.e., the factors important for capillarogenesis.

Keywords

Thalidomide Angiogenesis Cytochrome P-450 Apoptosis Caspase-3 
This is a preview of subscription content, log in to check access.

Notes

Acknowledgements

This work was partly supported by a grant to the Saitama Medical School Research Center for Genomic Medicine.

References

  1. Akita M, Murata E, Kaneko K, Ghaida J, Merker H-J (1993) Cell shape and arrangement of cultured aortic smooth muscle cells grown on collagen gels. Cell Tissue Res 274:91–95PubMedGoogle Scholar
  2. Akita M, Murata E, Merker H-J, Kaneko K (1997a) Formation of new capillary-like tubes in a three-dimensional in vitro model (aorta/collagen gel). Ann Anat 179:127–136Google Scholar
  3. Akita M, Murata E, Merker H-J, Kaneko K (1997b) Morphology of capillary-like structures in a three-dimensional aorta/collagen gel culture. Ann Anat 179:137–147Google Scholar
  4. Alnemri ES, Livingston DJ, Nicholson DW, Salvesen G, Thornberry NA, Wong WW, Yuan J (1996) Human ICE/CED-3 protease nomenclature. Cell 87:171PubMedGoogle Scholar
  5. Bauer KS, Dixon SC, Figg WD (1998) Inhibition of angiogenesis by thalidomide requires metabolic activation, which is species-dependent. Biochem Pharmacol 55:1827–1834CrossRefPubMedGoogle Scholar
  6. Bishop ET, Bell GT, Bloor S, Broom IJ, Hendy NF, Wheatley DN (1999) An in vitro model of angiogenesis: basic features. Angiogenesis 3:335–344CrossRefPubMedGoogle Scholar
  7. D’Amato RJ, Loughnan MS, Flynn E, Folkman J (1994) Thalidomide is an inhibitor of angiogenesis. Proc Natl Acad Sci U S A 91:4082–4085PubMedGoogle Scholar
  8. Diggle GE (2001) Thalidomide: 40 years on. Int J Clin Pract 55:627–631PubMedGoogle Scholar
  9. Dredge K, Marriott JB, Macdonald CD, Man H-W, Chen R, Muller GW, Stirling D, Dalgleish AG (2002) Novel thalidomide analogues display anti-angiogenic activity independently of immunomodulatory effects. Br J Cancer 87:1166–1172CrossRefPubMedGoogle Scholar
  10. Folkman J (1971) Tumor angiogenesis: therapeutic implications. N Engl J Med 181:182–186Google Scholar
  11. Fujita K, Asami Y, Murata E, Akita M, Kaneko K (2002) Effects of thalidomide, cytochrome P-450 and TNF-αon angiogenesis in a three-dimensional collagen gel-culture. Okajimas Folia Anat Jpn 79:101–106PubMedGoogle Scholar
  12. Han DC, Lee MY, Shin KD, Jeon SB, Kim JM, Son KH, Kim HC, Kim HM, Kwon BM (2004) 2′-benzoyloxycinnamaldehyde induces apoptosis in human carcinoma via reactive oxygen species. J Biol Chem (279:6911–6920). [Epub ahead of print]Google Scholar
  13. Lin HL, Liu TY, Wu CW, Chi CW (2001) 2-Methoxyestradiol-induced caspase-3 activation and apoptosis occurs through G (2)/M arrest dependent and independent pathways in gastric carcinoma cells. Cancer 92:500–509CrossRefPubMedGoogle Scholar
  14. Mitsiades N, Mitsiades CS, Poulaki V, Chauhan D, Richardson PG, Hideshima T, Munshi NC, Treon SP, Anderson KC (2002) Apoptotic signaling induced by immunomodulatory thalidomide analogs in human multiple myeloma cells: therapeutic implications. Blood 99:4525–4530CrossRefPubMedGoogle Scholar
  15. Nau H (1986) Species differences in pharmacokinetics and drug teratogenesis. Environ Health Perspect 70:113–129PubMedGoogle Scholar
  16. Neubert R, Merker H-J, Neubert D (1995) Developmental model for thalidomide action. Nature 400:1500–1502Google Scholar
  17. Parman T, Wiley MJ, Wells PG (1999) Free radical-medicated oxidative DNA damage in the mechanism of thalidomide teratogenicity. Nat Med 5:582–585CrossRefPubMedGoogle Scholar
  18. Rajkumar SV (2001) Thalidomide in the treatment of multiple myeloma. Expert Rev Anticancer Ther 1:2–28Google Scholar
  19. Rajkumar SV, Leong T, Roche PC, Fonseca R, Dispenzieri A, Lacy MQ, Lust JA, Witzig TE, Kyle RA, Gertz MA, Greipp PR (2000) Prognostic value of bone marrow angiogenesis in multiple myeloma. Clin Cancer Res 6:3111–3116PubMedGoogle Scholar
  20. Sauer H, Guenther J, Hescheler J, Wartenberg M (2000) Thalidomide inhibits angiogenesis in embryoid bodies by the generation of hydroxyl radicals. Am J Pathol 156:151–158PubMedGoogle Scholar
  21. Singhal S, Mehta J, Desikan R, Ayers D, Roberson P, Eddlemon P, Munshi N, Anaissie E, Wilson C, Dhodapkar M, Zeddis J, Barlogie B (1999) Antitumor activity of thalidomide in refractory multiple myeloma. N Engl J Med 341:1565–1571CrossRefPubMedGoogle Scholar
  22. Vacca A, Ribatti D, Roncali L, Ranieri G, Serio G, Silvestris F, Dammacco F (1994) Bone marrow angiogenesis and progression in multiple myeloma. Br J Haematol 87:503–508PubMedGoogle Scholar
  23. Vesela D, Vesely D, Jelinek R (1994) Embryotoxicity in chick embryo of thalidomide hydrolysis products following metabolic activation by rat liver homogenate. Funct Dev Morphol 4:313–316PubMedGoogle Scholar
  24. Wells PG, Kim PM, Laposa RR, Nicol CJ, Parman T, Winn L (1997) Oxidative damage in chemical teratogenesis. Mutat Res 396:65–78CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • Keiko Fujita
    • 1
  • Yoshiko Asami
    • 1
  • Kayoko Tanaka
    • 2
  • Masumi Akita
    • 2
  • Hans-Joachim Merker
    • 3
  1. 1.Department of AnatomySaitama Medical SchoolSaitamaJapan
  2. 2.Division of Morphological Science, Biomedical Research CenterSaitama Medical SchoolSaitamaJapan
  3. 3.Institute of AnatomyFree University of BerlinBerlinGermany

Personalised recommendations