Peroxisome proliferator-activated receptor-gamma activates p53 gene promoter binding to the nuclear factor-kappaB sequence in human MCF7 breast cancer cells.
The aim of the present study was to provide new mechanistic insight into the growth arrest and apoptosis elicited by peroxisome proliferator-activated receptor (PPAR)gamma in breast cancer cells. We ascertained that PPARgamma mediates the inhibition of cycle progression in MCF7 cells exerted by the specific PPARgamma agonist rosiglitazone [BRL4653 (BRL)], because this response was no longer notable in the presence of the receptor antagonist GW9662. We also provided evidence that BRL is able to up-regulate mRNA and protein levels of the tumor suppressor gene p53 and its effector p21(WAF1/Cip1) in a time- and dose-dependent manner. Moreover, in transfection experiments with deletion mutants of the p53 gene promoter, we documented that the nuclear factor-kappaB sequence is required for the transcriptional response to BRL. Interestingly, EMSA showed that PPARgamma binds directly to the nuclear factor-kappaB site located in the promoter region of p53, and chromatin immunoprecipitation experiments demonstrated that BRL increases the recruitment of PPARgamma on the p53 promoter sequence. Next, both PPARgamma and p53 were involved in the cleavage of caspases-9 and DNA fragmentation induced by BRL, given that GW9662 and an expression vector for p53 antisense blunted these effects. Our findings provide evidence that the PPARgamma agonist BRL promotes the growth arrest and apoptosis in MCF7 cells, at least in part, through a cross talk between p53 and PPARgamma, which may be considered an additional target for novel therapeutic interventions in breast cancer patients.
Bonofiglio
Daniela
D
Department of Pharmaco-Biology, University of Calabria, 87030 Arcavacata di Rende, Italy. daniela.bonofiglio@tin.it
Aquila
Saveria
S
Catalano
Stefania
S
Gabriele
Sabrina
S
Belmonte
Maria
M
Middea
Emilia
E
Qi
Hongyan
H
Morelli
Catia
C
Gentile
Mariaelena
M
Maggiolini
Marcello
M
Andò
Sebastiano
S
D020029:Base Pairing; D001943:Breast Neoplasms / Q000235:genetics / Q000378:metabolism*; D053453:Caspase 9 / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D047369:Chromatin Immunoprecipitation; D053938:DNA Fragmentation; D016193:G1 Phase / Q000187:drug effects; D016158:Genes, p53; D006801:Humans; D016328:NF-kappa B / Q000378:metabolism*; D047495:PPAR gamma / Q000819:agonists / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D016192:Resting Phase, Cell Cycle / Q000187:drug effects; D000077154:Rosiglitazone; D045162:Thiazolidinediones / Q000494:pharmacology; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism*
D016328:NF-kappa B; D047495:PPAR gamma; D045162:Thiazolidinediones; D016159:Tumor Suppressor Protein p53; D000077154:Rosiglitazone; D053453:Caspase 9