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J Biol Chem, Vol. 274, Issue 16, 11245-11252, April 16, 1999

The Role of Antiapoptotic Bcl-2 Family Members in Endothelial Apoptosis Elucidated with Antisense Oligonucleotides

Elizabeth J. Ackermann, Jennifer K. Taylor, Ranjit Narayana, and C. Frank Bennett

From the Department of Molecular Pharmacology, Isis Pharmaceuticals, Carlsbad, California 92008

In this study, we utilized potent antisense oligonucleotides to examine the role of two Bcl-2 family members found in human umbilical vein endothelial cells (HUVEC). The first, A1, is thought to be a TNF-alpha -inducible cytoprotective gene, and the second, Bcl-XL, is constitutively expressed.

Inhibition of the constitutive levels of Bcl-XL caused 10-25% of the cell population to undergo apoptosis and increased the susceptibility of cells to treatment with low concentrations of staurosporin or ceramide. The caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp(OMe)-CH2 prevented DNA fragmentation and Delta Ym loss caused by Bcl-XL inhibition or Bcl-XL inhibition combined with staurosporin. However, disruption of Delta Ym caused by Bcl-XL inhibition combined with ceramide treatment was not inhibited by benzyloxycarbonyl-Val-Ala-Asp(OMe)-CH2, although DNA fragmentation was completely prevented. Taken together, these results demonstrate a direct protective role for Bcl-XL under normal resting conditions and under low level apoptotic challenges to HUVEC. Furthermore, Bcl-XL protects cells from caspase-dependent and -independent mechanisms of Delta Ym disruption.

In contrast to Bcl-XL, A1 inhibition did not show a marked effect on the susceptibility of HUVEC to undergo apoptosis in response to TNF-alpha , ceramide, or staurosporin. These results demonstrate that although A1 may be a cytoprotective gene induced by TNF-alpha , it is not primarily responsible for HUVEC resistance to this cytokine.

Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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