title: Benzyl butyl phthalate induces necrosis by AhR mediation of CYP1B1 expression in human granulosa cells.
abstract: We investigated the signaling pathway of the arylhydrocarbon receptor (AhR) on HO23 cells (immortalized human granulosa cells (hGC)) mediated by benzyl butyl-phthalate (BBP). BBP (1 μM) significantly increased the mRNA and protein levels of AhR, aryl hydrocarbon receptor nuclear translocator (ARNT) and cytochrome-P450 (CYP)1B1 in HO23 cells. Treatment with 3',4'-dimethoxyflavone (3',4'-DMF) or AhR siRNA significantly reduced AhR and CYP1B1, but CYP1A1 was not affected by 3',4'-DMF or AhR siRNA, suggesting that increases in CYP1A1 may not regulated by AhR. BBP induced the AhR fusion protein to localize and accumulate around the nucleus, and AhR heterodimerization with ARNT was observed in the nucleus by immunoprecipitation. Chromatin immunoprecipitation and reporter assays revealed the effect of BBP on CYP1B1, but not CYP1A1. Necrosis was significantly increased in HO23 cells after BBP treatment, and 3',4'-DMF, AhR siRNA or CYP1B1 siRNA knockdown blocked this phenomenon. These data suggest that BBP-induced HO23 cell necrosis is AhR and CYP1B1 dependent.
authors.affiliation: Department of Obstetrics and Gynecology, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan.
authors.forename: Hung-Sheng
authors.initials: HS
authors.lastname: Chen
authors.forename: Po-Hui
authors.initials: PH
authors.lastname: Chiang
authors.forename: Yu-Chih
authors.initials: YC
authors.lastname: Wang
authors.forename: Mi-Chun
authors.initials: MC
authors.lastname: Kao
authors.forename: Tsung-Hua
authors.initials: TH
authors.lastname: Shieh
authors.forename: Cheng-Fang
authors.initials: CF
authors.lastname: Tsai
authors.forename: Eing-Mei
authors.initials: EM
authors.lastname: Tsai
mesh_terms: D021581:Active Transport, Cell Nucleus; D017209:Apoptosis / Q000187:drug effects; D001189:Aryl Hydrocarbon Hydroxylases / Q000235:genetics / Q000378:metabolism*; D051784:Aryl Hydrocarbon Receptor Nuclear Translocator / Q000235:genetics / Q000378:metabolism; D051792:Basic Helix-Loop-Helix Proteins / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D047369:Chromatin Immunoprecipitation; D019363:Cytochrome P-450 CYP1A1 / Q000235:genetics / Q000378:metabolism; D065633:Cytochrome P-450 CYP1B1; D004305:Dose-Response Relationship, Drug; D005260:Female; D047309:Flavones / Q000494:pharmacology; D017930:Genes, Reporter; D006107:Granulosa Cells / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D006801:Humans; D009336:Necrosis; D010795:Phthalic Acids / Q000633:toxicity*; D010968:Plasticizers / Q000633:toxicity*; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D018336:Receptors, Aryl Hydrocarbon / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D013997:Time Factors; D014162:Transfection; D015854:Up-Regulation
chemical_list: D001189:Aryl Hydrocarbon Hydroxylases; D051784:Aryl Hydrocarbon Receptor Nuclear Translocator; D051792:Basic Helix-Loop-Helix Proteins; D019363:Cytochrome P-450 CYP1A1; D065633:Cytochrome P-450 CYP1B1; D047309:Flavones; D010795:Phthalic Acids; D010968:Plasticizers; D012333:RNA, Messenger; D018336:Receptors, Aryl Hydrocarbon; C509288:3',4'-dimethoxyflavone; C111235:AHR protein, human; C497396:ARNT protein, human; C569463:CYP1A1 protein, human; C585496:CYP1B1 protein, human; C027561:butylbenzyl phthalate