Abstract
Background: Butyrate, a potent histone deacetylase inhibitor, belongs to a promising new class of antineoplastic agents with the capacity to induce apoptosis of cancer cells. However, the underlying mechanisms of action have yet not been elucidated. Aim: To further investigate the molecular events involved in butyrate-induced caspase-3 activation in Caco-2 wild-type, empty-vector and dominant-negative PPARγ mutant cells along the signalling pathway. In this context, the involvement and up-regulation of PPARγ was examined. Results: Stimulation of cells with butyrate resulted in increased expression of PPARγ mRNA, protein, and activity as well as phospho-p38 MAPK protein expression and caspase-3 activity. Arsenite, a direct stimulator of p38 MAPK, also led to an increased PPARγ expression, thereby mimicking the effects of butyrate. In contrast, butyrate-mediated up-regulation of PPARγ was counteracted by co-incubation with the p38 MAPK inhibitor SB203580. Treatment of cells with butyrate resulted in both increased caspase-8 and -9 activity and reduced expression of XIAP and survivin. However, butyrate-mediated effects on these apoptosis-regulatory proteins leading to caspase-3 activation were almost completely abolished in Caco-2 dominant-negative PPARγ mutant cells. Conclusions: Our data clearly unveil PPARγ as a key target in the butyrate-induced signalling cascade leading to apoptosis via caspase-3 in Caco-2 cells.
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Abbreviations
- DFF:
-
DNA Fragmentation Factor
- DMEM:
-
Dulbecco’s modified Eagle’s medium
- DMSO:
-
Dimethyl sulfoxide
- ECACC:
-
European Collection of cell cultures
- EGF:
-
Epithelial growth factor
- ERK:
-
Extracellular signal-regulated kinase
- FCS:
-
Foetal calf serum
- FLIP:
-
FLICE-inhibitory protein
- HDAC:
-
Histone deacetylase
- IAP:
-
Inhibitor of apoptosis protein
- JNK:
-
cJun N-terminal kinase
- LDH:
-
Lactate dehydrogenase
- MAPK:
-
Mitogen-activated protein kinase
- PARP:
-
Poly(ADP-ribose) Polymerase
- PBS:
-
Phosphate buffered saline
- PKC-δ:
-
Protein kinase C-δ
- PPARγ:
-
Peroxisome proliferator-activated receptor γ
- SCFA:
-
Short chain fatty acid
- STAT3:
-
Signal transducer and activator of transcription-3
- TBS-T:
-
Tris-buffered saline containing 0.05% Tween 20
- TNF:
-
Tumor necrosis factor
- XIAP:
-
X linked inhibitor of apoptosis
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Schwab, M., Reynders, V., Ulrich, S. et al. PPARγ is a key target of butyrate-induced caspase-3 activation in the colorectal cancer cell line Caco-2. Apoptosis 11, 1801–1811 (2006). https://doi.org/10.1007/s10495-006-9788-2
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DOI: https://doi.org/10.1007/s10495-006-9788-2