Endostatin Induces Endothelial Cell Apoptosis*
- Mohanraj Dhanabal‡§,
- Ramani Ramchandran‡,
- Matthew J. F. Waterman§,
- Hua Lu,
- Bertrand Knebelmann,
- Mark Segal and
- Vikas P. Sukhatme¶
- From the Renal Division, Department of Medicine, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, Massachusetts 02215
Abstract
Endostatin, a carboxyl-terminal fragment of collagen XVIII, has been shown to regress tumors in mice. In this study, we have analyzed the mechanism of endostatin action on endothelial cells and nonendothelial cells. Endostatin treatment of cow pulmonary artery endothelial cells caused apoptosis, as demonstrated by three methods, annexin V-fluorescein isothiocyanate staining, caspase 3, and terminal deoxynucleotidyl transferase-mediated dUTP nick-end-labeling assay. Moreover, addition of endostatin led to a marked reduction of the Bcl-2 and Bcl-XLanti-apoptotic protein, whereas Bax protein levels were unaffected. These effects were not seen in several nonendothelial cells. Collectively, these findings provide important mechanistic insight into endostatin action.
Footnotes
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↵* This work was supported by seed funds from Beth Israel Deaconess Medical Center.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
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↵§ Recipients of salary support from a National Institutes of Health basic science training grant.
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↵‡ These authors made equal contributions to this work.
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↵¶ To whom correspondence should be addressed: Beth Israel Deaconess Medical Center, Dana 517, 330 Brookline Ave., Boston, MA 02215. Tel.: 617-667-2105; Fax: 617-667-7843; E-mail:vsukhatm{at}bidmc.harvard.edu.
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↵2 M. Dhanabal, R. Volk, R. Ramchandran, M. Simons, and V. P. Sukhatme, manuscript in preparation.
- Abbreviations:
- FITC
-
fluorescein isothiocyanate
- FACS
-
fluorescence-activated cell sorting
- TUNEL
-
terminal deoxynucleotidyl transferase-mediated dUTP nick-end-labeling
- TNF
-
tumor necrosis factor
- DMEM
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Dulbecco’s modified Eagle’s medium
- C-PAE
-
cow pulmonary artery endothelial
- PBS
-
phosphate-buffered saline
- PI
-
propidium iodide
- FGF
-
fibroblast growth factor
- PS
-
phosphatidylserine
- FCS
-
fetal calf serum
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- Received October 14, 1998.
- Revision received December 17, 1998.
- The American Society for Biochemistry and Molecular Biology, Inc.
