title: Peroxisome proliferator-activated receptor gamma (PPARgamma) regulates trefoil factor family 2 (TFF2) expression in gastric epithelial cells.
abstract: Although trefoil factor family 2 (TFF2) plays a critical role in the defense and repair of gastric mucosa, the regulatory mechanism of TFF2 expression is not fully understood. In this study, we investigated the regulation of TFF2 expression by peroxisome proliferator-activated receptor gamma (PPARgamma) in gastric epithelial cells. MKN45 gastric cells were used. TFF2 mRNA expression was analyzed by real-time quantitative RT-PCR. The promoter sequence of the human TFF2 gene was cloned into pGL3-basic vector for reporter gene assays. Ciglitazone was mainly used as a specific PPARgamma ligand. MKN45 cells expressed functional PPARgamma proteins. Endogenous TFF2 mRNA expression and TFF2 reporter gene transcription was significantly up-regulated by ciglitazone in a dose-dependent manner. Reporter gene assays showed that two distinct cis-elements are involved in the response to PAPRgamma activation. Within one of these element (nucleotides -558 to -507), we identified a functional peroxisome proliferator responsive element (PPRE) at -522 (5'-GGGACAAAGGGCA-3'). Electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP) assay confirmed the binding of PPARgamma to this sequence. Another element (nucleotides -407 to -358) appeared to be a composite enhancer element indirectly regulated by PPARgamma and a combination of these two cis-elements was required for the full response of the human TFF2 gene expression to PPARgamma. These data demonstrate that human TFF2 gene is a direct target of PPARgamma in gastric epithelial cells. Since TFF2 is a critical gastroprotective agent, PPARgamma may be involved in the gastric mucosal defense through regulating TFF2 expression in humans.
authors.affiliation: Department of Gastroenterology, Dokkyo Medical University, Mibu, Tochigi 321-0293, Japan. tshimada@dokkyomed.ac.jp
authors.forename: Tadahito
authors.initials: T
authors.lastname: Shimada
authors.forename: Yoichiro
authors.initials: Y
authors.lastname: Fujii
authors.forename: Takero
authors.initials: T
authors.lastname: Koike
authors.forename: Kyoko
authors.initials: K
authors.lastname: Tabei
authors.forename: Takashi
authors.initials: T
authors.lastname: Namatame
authors.forename: Michiko
authors.initials: M
authors.lastname: Yamagata
authors.forename: Akihiro
authors.initials: A
authors.lastname: Tajima
authors.forename: Masashi
authors.initials: M
authors.lastname: Yoneda
authors.forename: Akira
authors.initials: A
authors.lastname: Terano
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Hiraishi
mesh_terms: D001483:Base Sequence; D002460:Cell Line; D047369:Chromatin Immunoprecipitation; D017931:DNA Primers / Q000235:genetics; D024202:Electrophoretic Mobility Shift Assay; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D005753:Gastric Mucosa / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D047495:PPAR gamma / Q000819:agonists / Q000235:genetics / Q000378:metabolism*; D010455:Peptides / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D045162:Thiazolidinediones / Q000494:pharmacology; D000071164:Trefoil Factor-2
chemical_list: D017931:DNA Primers; D047495:PPAR gamma; D010455:Peptides; D012333:RNA, Messenger; C000604007:TFF2 protein, human; D045162:Thiazolidinediones; D000071164:Trefoil Factor-2; C039671:ciglitazone