The peroxisome proliferator BR931 kills FaO cells by p53-dependent apoptosis.
Although suppression of apoptosis has been implicated as a mechanism for the hepatocarcinogenicity of peroxisome proliferators (PPs), they can also induce cell death in rat AH130 and human HepG2 hepatoma cells. To study how PPs induce cell death and to characterize the molecular events involved, we administered the hypolipidemic BR931, a peroxisome proliferator, to rat hepatoma FaO cells. Treatment with increasing concentrations of BR931 (0.015 to 0.6 mM) reduced cell viability in a dose- and time-dependent manner, associated with DNA fragmentation and morphological changes characteristic of apoptosis. BR931 also caused phosphorylation of p53 within 3 hours, translocation of the pro-apoptotic Bax protein to mitochondria, release of cytochrome-c into the cytosol, and activation of caspase-9 and -3. These results indicated that BR931 activated the intrinsic caspase cascade. Pretreatment with three different antioxidants, N-acetylcysteine, Vitamin C and Trolox, reduced apoptosis, suggesting that reactive oxygen species (ROS) plays a role in BR931-induced apoptosis. In support of this hypothesis, BR931 produced increased levels of 8-hydroxy-deoxy-guanosine, a marker of DNA oxidative damage. Antioxidants prevented the p53 phosphorylation, up-regulation of Bax and BR931-induced apoptosis. These results suggest that BR931 can increase generation of ROS, leading to DNA damage and p53 phosphorylation, which, in turn, induces the activation of Bax, release of cytochrome-c from mitochondria and activation of caspases, culminating in cell death.
Simbula
Gabriella
G
Department of Toxicology, Unit of Oncology and Molecular Pathology, Via Porcell 4, 09124 Cagliari, Italy. gmledda@unica.it
Pibiri
Monica
M
Sanna
Laura
L
Cossu
Costanza
C
Molotzu
Francesca
F
Columbano
Amedeo
A
Ledda-Columbano
Giovanna M
GM
D000111:Acetylcysteine / Q000378:metabolism; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D017209:Apoptosis / Q000235:genetics / Q000502:physiology*; D001205:Ascorbic Acid / Q000494:pharmacology; D015153:Blotting, Western; D053148:Caspase 3; D053453:Caspase 9; D020169:Caspases / Q000378:metabolism; D045744:Cell Line, Tumor; D002470:Cell Survival; D045304:Cytochromes c / Q000502:physiology; D003600:Cytosol / Q000378:metabolism; D004247:DNA / Q000737:chemistry; D053938:DNA Fragmentation / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D004587:Electrophoresis, Agar Gel; D016158:Genes, p53 / Q000502:physiology*; D006801:Humans; D020287:In Situ Nick-End Labeling; D008114:Liver Neoplasms, Experimental / Q000473:pathology; D008928:Mitochondria / Q000378:metabolism; D020025:Peroxisome Proliferators / Q000494:pharmacology*; D010766:Phosphorylation; D021381:Protein Transport / Q000502:physiology; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000502:physiology; D019253:Proto-Oncogene Proteins c-bcl-2; D011743:Pyrimidines / Q000494:pharmacology*; D051381:Rats; D013997:Time Factors; D051028:bcl-2-Associated X Protein
D000975:Antioxidants; C494826:BAX protein, human; C494828:Bax protein, rat; D020025:Peroxisome Proliferators; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D011743:Pyrimidines; D051028:bcl-2-Associated X Protein; D045304:Cytochromes c; D004247:DNA; C016908:pirinixil; C505407:CASP3 protein, human; C506433:CASP9 protein, human; C505410:Casp3 protein, rat; C506435:Casp9 protein, rat; D053148:Caspase 3; D053453:Caspase 9; D020169:Caspases; D001205:Ascorbic Acid; D000111:Acetylcysteine