title: Induction of PUMA-alpha and down-regulation of PUMA-beta expression is associated with benzo(a)pyrene-induced apoptosis in MCF-7 cells.
abstract: Benzo(a)pyrene (BP) forms benzo(a)pyrene-7,8-dihydrodiol-9,10-epoxide (BPDE)-DNA adducts in human breast adenocarcinoma MCF-7 cells, leading to p53 protein induction and phosphorylation. Although BP-induced apoptosis in rodent cells is known, it is still unclear in human cells. Here we have analyzed the effects of BP on p53 related apoptotic proteins, cell cycle and cell death in MCF-7 cells. PUMA-protein (p53 up-regulated modulator of apoptosis) levels were changed after BP exposure so that PUMA-alpha protein was statistically significantly increased whereas PUMA-beta protein was statistically significantly decreased. PUMA-protein levels were also investigated in ZR-75-1 cells, where PUMA-alpha protein was statistically significantly increased. Cytochrome c, which is released from mitochondria during apoptosis to form the apoptosome, was increased in cytoplasmic fraction after BP exposure in MCF-7 cells. Increased apoptosis was also seen after 48 and 72 h BP exposure (2.5 and 5 microM). In addition, BP decreased dose dependently cell viability (2.5 and 5 microM) and increased ROS formation (1 and 10 microM). Our results suggest that PUMA-alpha protein is involved in BP-induced cell death most likely through a p53 dependent apoptotic pathway.
authors.affiliation: University of Kuopio, Department of Pharmacology and Toxicology, Yliopistonranta 1C, Kuopio, Finland. Marjo.Tampio@uku.fi
authors.forename: Marjo
authors.initials: M
authors.lastname: Tampio
authors.forename: Piia
authors.initials: P
authors.lastname: Markkanen
authors.forename: Katja A
authors.initials: KA
authors.lastname: Puttonen
authors.forename: Eveliina
authors.initials: E
authors.lastname: Hagelberg
authors.forename: Hannu
authors.initials: H
authors.lastname: Heikkinen
authors.forename: Kati
authors.initials: K
authors.lastname: Huhtinen
authors.forename: Jarkko
authors.initials: J
authors.lastname: Loikkanen
authors.forename: Maija-Riitta
authors.initials: MR
authors.lastname: Hirvonen
authors.forename: Kirsi H
authors.initials: KH
authors.lastname: Vähäkangas
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D051017:Apoptosis Regulatory Proteins / Q000096:biosynthesis*; D001564:Benzo(a)pyrene / Q000633:toxicity*; D045744:Cell Line, Tumor; D045304:Cytochromes c / Q000378:metabolism; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism; D015536:Down-Regulation; D005434:Flow Cytometry; D006801:Humans; D015151:Immunoblotting; D008564:Membrane Potentials / Q000187:drug effects; D018613:Microscopy, Confocal; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D020033:Protein Isoforms; D011518:Proto-Oncogene Proteins / Q000096:biosynthesis*; D017382:Reactive Oxygen Species / Q000378:metabolism; D015854:Up-Regulation
chemical_list: D051017:Apoptosis Regulatory Proteins; C438378:BBC3 protein, human; D020033:Protein Isoforms; D011518:Proto-Oncogene Proteins; D017382:Reactive Oxygen Species; D001564:Benzo(a)pyrene; D045304:Cytochromes c