Caspases-2, -3, and -7 are involved in thapsigargin-induced apoptosis of SH-SY5Y neuroblastoma cells.

Abstract

Caspase-2 has been reported to play a role in the cell death observed under a number of different conditions; however, it is unclear whether caspase-2 plays a role in cell death triggered by endoplasmic reticulum (ER) stress. The purpose of this study was to determine whether caspase-2 is involved in SH-SY5Y neuroblastoma cell death caused by thapsigargin-induced ER stress. Thapsigargin treatment (1 microM, 16 hr) stimulated the proteolytic processing of caspases-2, -3, and -7, suggesting that these caspases are activated by ER stress. The role of these caspases in thapsigargin-induced cell death was examined by using cell-permeable caspase inhibitors. In the absence of pretreatment with caspase inhibitors, thapsigargin (0.1 microM, 20 hr) reduced the number of viable cells to 53.9% +/- 3.3% of starting-time control. Pretreatment for 90 min with either the pan-caspase inhibitor Z-VAD-FMK or the caspase-2-selective inhibitor Z-VDVAD-FMK inhibited thapsigargin-stimulated cell death, resulting in the number of viable cells being 115.6% +/- 5.3% (P < 0.001) and 69.3% +/- 2.9% (P < 0.01), respectively, of starting-time control. Neither the caspase-3- and -7-selective inhibitor Z-DEVD-FMK nor the caspase-9-selective inhibitor Z-LEHD-FMK significantly affected thapsigargin-stimulated cell death. An anticaspase-12-reactive protein was also identified in SH-SY5Y cells, but thapsigargin had no effect on proteolysis of this protein. These data demonstrate that caspases-2, -3, and -7 are involved in ER stress-mediated death of SH-SY5Y cells.

• Authors: Mary K Dahmer
• Journal: Journal of neuroscience research (J Neurosci Res) Vol. 80 Issue 4 Pg. 576-83 (May 15 2005) ISSN: 0360-4012 [Print] United States
• PMID: 15825194 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Caspase Inhibitors
• Enzyme Inhibitors
• Thapsigargin
• CASP3 protein, human
• CASP7 protein, human
• Caspase 2
• Caspase 3
• Caspase 7
• Caspases

Topics

• Analysis of Variance
• Apoptosis (drug effects)
• Blotting, Western (methods)
• Caspase 2
• Caspase 3
• Caspase 7
• Caspase Inhibitors
• Caspases (metabolism)
• Cell Line, Tumor
• Drug Interactions
• Enzyme Inhibitors (toxicity)
• Humans
• Neuroblastoma
• Thapsigargin (toxicity)