Multiple forms of angiostatin induce apoptosis in endothelial cells

Rudolf Lucas, L. Holmgren, I. Garcia, B. Jimenez, S. J. Mandriota, F. Borlat, B. K L Sim, Z. Wu, G. E. Grau, Y. Shing, G. A. Soff, N. Bouck, M. S. Pepper

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207 Scopus citations

Abstract

Angiostatin is a circulating inhibitor of angiogenesis generated by proteolytic cleavage of plasminogen. In this study We have used recombinant human and murine angiostatins (kringles 1-4) as well as native human angiostatin (prepared by elastase digestion of plasminogen [kringles 1-3] or by plasmin autocatalysis in the presence of a free sulfhydryl donor [kringles 1-4]). We report that angiostatin reduces endothelial cell number in a 4-day proliferation assay without affecting cell cycle progression into S-phase (as determined by bromodeoxyuridine labeling). This suggested that the reduction in cell number in the proliferation assay might in part be due to cytotoxicity. This was confirmed by the observation that ethidium homodimer incorporation (a measure of plasma membrane integrity) into endothelial cells was increased by angiostatin in a manner similar to that seen with tumor necrosis factor-α (TNF-α) and transforming growth factor-β1 (TGF-β1), both of which induce apoptosis in endothelial cells. In contrast to TNF-α and TGF-β1, angiostatin did not induce cytotoxicity in human MRC-5 fibroblast, rat smooth muscle, canine MDCK epithelial, or murine B16-F10 melanoma cell lines. Angiostatin-induced apoptosis was confirmed by endothelial cell nuclear acridine orange incorporation as well as by annexin V and TUNEL staining. These in vitro findings point to endothelial cell apoptosis as a mechanism for the antiangiogenic effect of angiostatin in vivo.

Original languageEnglish (US)
Pages (from-to)4730-4741
Number of pages12
JournalBlood
Volume92
Issue number12
StatePublished - Dec 15 1998
Externally publishedYes

ASJC Scopus subject areas

  • Immunology
  • Biochemistry
  • Hematology
  • Cell Biology

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    Lucas, R., Holmgren, L., Garcia, I., Jimenez, B., Mandriota, S. J., Borlat, F., Sim, B. K. L., Wu, Z., Grau, G. E., Shing, Y., Soff, G. A., Bouck, N., & Pepper, M. S. (1998). Multiple forms of angiostatin induce apoptosis in endothelial cells. Blood, 92(12), 4730-4741.