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PPARγ is a key target of butyrate-induced caspase-3 activation in the colorectal cancer cell line Caco-2

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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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Correspondence to Oliver Schröder.

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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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