A dominant negative form of IKK2 prevents suppression of apoptosis by the peroxisome proliferator nafenopin.
Peroxisome proliferators (PPs) are a class of non-genotoxic chemicals that cause rodent liver enlargement and hepatocarcinogenesis. In primary rat hepatocyte cultures, PPs suppress spontaneous apoptosis and that induced by a number of pro-apoptotic stimuli such as transforming growth factor-beta(1). Tumour necrosis factor alpha (TNF-alpha) and the transcription factor NFkappaB have been implicated in the mode of action of PPs. TNF-alpha signalling to NFkappaB is thought to be responsible for many of the effects elicited by this cytokine. NFkappaB regulates gene expression in immunity, stress responses and the inhibition of apoptosis. Activation of NFkappaB requires the successive action of NFkappaB-inducing kinase and the phosphorylation of NFkappaB inhibitory proteins (IkappaB) by an IkappaB kinase (IKK) complex. The IKK2 subunit of IkappaB kinase is thought to be essential for NFkappaB activation and prevention of apoptosis. To determine whether IKK2 plays a role in the suppression of apoptosis by PPs, we expressed a dominant negative form of IKK2 (IKK2dn) in primary rat hepatocyte cultures. Infection with an adenovirus construct expressing IKK2dn caused apoptosis in control primary rat hepatocytes in the absence of exogenous TNF-alpha. Moreover, IKK2dn-induced apoptosis could not be rescued by addition of TNF-alpha or the peroxisome proliferator nafenopin. These results demonstrate a requirement for intracellular signalling pathways mediated by IKK2 in the suppression of apoptosis by the PP class of hepatocarcinogens.
Cosulich
S C
SC
AstraZeneca Pharmaceuticals and Zeneca Central Toxicology Laboratory, 3G8 Mereside, Alderley Park, Macclesfield, Cheshire SK10 4TG, UK. sabina.cosulich@astrazeneca.com
James
N H
NH
Needham
M R
MR
Newham
P P
PP
Bundell
K R
KR
Roberts
R A
RA
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D005786:Gene Expression Regulation; D051550:I-kappa B Kinase; D051034:Inhibitor of Apoptosis Proteins; D008099:Liver / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D016328:NF-kappa B / Q000502:physiology; D009255:Nafenopin / Q000037:antagonists & inhibitors / Q000633:toxicity; D020025:Peroxisome Proliferators / Q000037:antagonists & inhibitors / Q000633:toxicity; D014176:Protein Biosynthesis; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000502:physiology*; D011506:Proteins / Q000235:genetics; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D014161:Transduction, Genetic; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology / Q000502:physiology
D051034:Inhibitor of Apoptosis Proteins; D016328:NF-kappa B; D020025:Peroxisome Proliferators; D011506:Proteins; D014409:Tumor Necrosis Factor-alpha; D009255:Nafenopin; D017346:Protein Serine-Threonine Kinases; D051550:I-kappa B Kinase