RRC ID 45255
著者 Kageyama-Yahara N, Yamamichi N, Takahashi Y, Nakayama C, Shiogama K, Inada K, Konno-Shimizu M, Kodashima S, Fujishiro M, Tsutsumi Y, Ichinose M, Koike K.
タイトル Gli regulates MUC5AC transcription in human gastrointestinal cells.
ジャーナル PLoS One
Abstract MUC5AC is a well-known gastric differentiation marker, which has been frequently used for the classification of stomach cancer. Immunohistochemistry revealed that expression of MUC5AC decreases accompanied with increased malignant property of gastric mucosa, which further suggests the importance of MUC5AC gene regulation. Alignment of the 5'-flanking regions of MUC5AC gene of 13 mammal species denoted high homology within 200 bp upstream of the coding region. Luciferase activities of the deletion constructs containing upstream 451 bp or shorter fragments demonstrated that 15 bp region between -111 and -125 bp plays a critical role on MUC5AC promoter activity in gastrointestinal cells. We found a putative Gli-binding site in this 15 bp sequence, and named this region a highly conserved region containing a Gli-binding site (HCR-Gli). Overexpression of Gli homologs (Gli1, Gli2, and Gli3) clearly enhanced MUC5AC promoter activity. Exogenous modulation of Gli1 and Gli2 also affected the endogenous MUC5AC gene expression in gastrointestinal cells. Chromatin immunoprecipitation assays demonstrated that Gli1 directly binds to HCR-Gli: Gli regulates MUC5AC transcription via direct protein-DNA interaction. Conversely, in the 30 human cancer cell lines and various normal tissues, expression patterns of MUC5AC and Gli did not coincide wholly: MUC5AC showed cell line-specific or tissue-specific expression whereas Gli mostly revealed ubiquitous expression. Luciferase promoter assays suggested that the far distal MUC5AC promoter region containing upstream 4010 bp seems to have several enhancer elements for gene transcription. In addition, treatments with DNA demethylation reagent and/or histone deacetylase inhibitor induced MUC5AC expression in several cell lines that were deficient in MUC5AC expression. These results indicated that Gli is necessary but not sufficient for MUC5AC expression: namely, the multiple regulatory mechanisms should work in the distal promoter region of MUC5AC gene.
巻・号 9(8)
ページ e106106
公開日 2014-1-1
DOI 10.1371/journal.pone.0106106
PII PONE-D-14-24645
PMID 25166306
PMC PMC4148389
MeSH Base Sequence Binding Sites Cell Line, Tumor Conserved Sequence Gastrointestinal Neoplasms / genetics Gastrointestinal Neoplasms / metabolism Gastrointestinal Neoplasms / pathology* Gastrointestinal Tract / metabolism* Gastrointestinal Tract / pathology Gene Expression Regulation, Neoplastic Humans Mucin 5AC / chemistry Mucin 5AC / genetics* Mucin 5AC / metabolism* Organ Specificity Promoter Regions, Genetic* Sequence Alignment Transcription Factors / genetics Transcription Factors / metabolism* Zinc Finger Protein GLI1
IF 2.74
引用数 8
WOS 分野 MULTIDISCIPLINARY SCIENCES
リソース情報
ヒト・動物細胞 T98G(RCB1954) A172(RCB2530)