Characterization of Apt- cell lines exhibiting crossresistance to glucocorticoid- and Fas-mediated apoptosis

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1999_Askew_Characterization.pdf
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1999
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Askew, David
Kuscuoglu, Unsal
Green, Douglas R.
Miesfeld, Roger L.
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Apoptosis induction by staurosporine, ceramide, and Fas stimulation was investigated in the mouse thymoma cell line W7.2 and a panel of dexamethasone (dex)-resistant W7.2 mutant cell lines, Apt3.8, Apt4.8 and Apt5.8, and a Bcl-2 transfectedW7.2 cell line (Wbcl2).WhileW7.2 cellswere found to be sensitive to these apoptosis inducers, the Apt- mutants andWbcl2 cells were shownto be resistant tosome or all of the treatments.Specifically, all threeApt-mutantsandWbcl2cells were found to be resistant to ceramide and Fas-mediated apoptosis, whereas, Apt4.8 and Apt5.8 were sensitive to staurosporine-induced apoptosis under conditions in which Apt3.8 and Wbcl2 cells were resistant. Measurements of caspase activity and cytochrome c release in cytosolic extracts of dex and staurosporine-treated cells indicated that the recessive Apt- mutations effect steps upstream of mitochondrial dysfunction. Steady-state RNA levels of apoptosis-associated gene transcripts showed that the observed differential resistance of the Apt- cell lines could not be explained by altered expression of numerous Bcl-2 or Fas related genes. Transient transfection of human Fas gene coding sequences into the Apt- mutants and Wbcl2 cells did not induce apoptosis, even though these same cell lines were sensitive to ectopic expression of the FADD and caspase 8 genes. Taken together, these data provide genetic evidence for the existence of shared components in the dex- and Fasmediated apoptotic pathways in W7.2 cells.

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570 Biowissenschaften, Biologie
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apoptosis, thymocytes, glucocorticoids, Fas, Bcl-2, staurosporine
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ISO 690ASKEW, David, Unsal KUSCUOGLU, Thomas BRUNNER, Douglas R. GREEN, Roger L. MIESFELD, 1999. Characterization of Apt- cell lines exhibiting crossresistance to glucocorticoid- and Fas-mediated apoptosis. In: Cell Death and Differentiation. 1999, 6(8), pp. 796-804
BibTex
@article{Askew1999Chara-16550,
  year={1999},
  title={Characterization of Apt- cell lines exhibiting crossresistance to glucocorticoid- and Fas-mediated apoptosis},
  number={8},
  volume={6},
  journal={Cell Death and Differentiation},
  pages={796--804},
  author={Askew, David and Kuscuoglu, Unsal and Brunner, Thomas and Green, Douglas R. and Miesfeld, Roger L.}
}
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    <dcterms:abstract xml:lang="eng">Apoptosis induction by staurosporine, ceramide, and Fas stimulation was investigated in the mouse thymoma cell line W7.2 and a panel of dexamethasone (dex)-resistant W7.2 mutant cell lines, Apt3.8, Apt4.8 and Apt5.8, and a Bcl-2 transfectedW7.2 cell line (Wbcl2).WhileW7.2 cellswere found to be sensitive to these apoptosis inducers, the Apt- mutants andWbcl2 cells were shownto be resistant tosome or all of the treatments.Specifically, all threeApt-mutantsandWbcl2cells were found to be resistant to ceramide and Fas-mediated apoptosis, whereas, Apt4.8 and Apt5.8 were sensitive to staurosporine-induced apoptosis under conditions in which Apt3.8 and Wbcl2 cells were resistant. Measurements of caspase activity and cytochrome c release in cytosolic extracts of dex and staurosporine-treated cells indicated that the recessive Apt- mutations effect steps upstream of mitochondrial dysfunction. Steady-state RNA levels of apoptosis-associated gene transcripts showed that the observed differential resistance of the Apt- cell lines could not be explained by altered expression of numerous Bcl-2 or Fas related genes. Transient transfection of human Fas gene coding sequences into the Apt- mutants and Wbcl2 cells did not induce apoptosis, even though these same cell lines were sensitive to ectopic expression of the FADD and caspase 8 genes. Taken together, these data provide genetic evidence for the existence of shared components in the dex- and Fasmediated apoptotic pathways in W7.2 cells.</dcterms:abstract>
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