Abstract
There is increasing evidence that proteases other than caspases, for example, the lysosomal cathepsins B, D and L, are involved in apoptotic cell death. In the present study, we present data that suggest a role for cathepsin D in staurosporine-induced apoptosis in human foreskin fibroblasts. Cathepsin D and cytochrome c were detected partially released to the cytosol after exposure to 0.1 μM staurosporine for 1 h. After 4 h, activation of caspase-9 and -3 was initiated and later caspase-8 activation and a decrease in full-length Bid were detected. Pretreatment of cells with the cathepsin D inhibitor, pepstatin A, prevented cytochrome c release and caspase activation, and delayed cell death. These results imply that cytosolic cathepsin D is a key mediator in staurosporine-induced apoptosis. Analysis of the relative sequence of apoptotic events indicates that, in this cell type, cathepsin D acts upstream of cytochrome c release and caspase activation.
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Abbreviations
- AMC:
-
7-amino-4-methyl coumarin
- AFC:
-
7-amino-4-trifluoromethyl coumarin
- DPPD:
-
N,N′-diphenyl-1,4-phenylene-diamine
- GAPDH:
-
glyceraldehyde-3-phosphate dehydrogenase
- PBS:
-
phosphate-buffered saline
- TBS:
-
Tris-buffered saline
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Acknowledgements
This work was financially supported by the Swedish Cancer Foundation (Grant No. 2703), the Swedish Research Council (Grant No. 14259) and Östergötlands Iäns landsting. We thank Dr. Katarina Kägedal and Dr. Uno Johansson for fruitful discussions and Dr. Brett Garner for linguistic help.
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Johansson, AC., Steen, H., Öllinger, K. et al. Cathepsin D mediates cytochrome c release and caspase activation in human fibroblast apoptosis induced by staurosporine. Cell Death Differ 10, 1253–1259 (2003). https://doi.org/10.1038/sj.cdd.4401290
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DOI: https://doi.org/10.1038/sj.cdd.4401290
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