Merck

Gli regulates MUC5AC transcription in human gastrointestinal cells.

PloS one (2014-08-29)
Natsuko Kageyama-Yahara, Nobutake Yamamichi, Yu Takahashi, Chiemi Nakayama, Kazuya Shiogama, Ken-ichi Inada, Maki Konno-Shimizu, Shinya Kodashima, Mitsuhiro Fujishiro, Yutaka Tsutsumi, Masao Ichinose, Kazuhiko Koike
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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
苯酚 溶液, Equilibrated with 10 mM Tris HCl, pH 8.0, 1 mM EDTA, BioReagent, for molecular biology
Sigma-Aldrich
苯酚, puriss. p.a., ACS reagent, reag. Ph. Eur., 99.0-100.5%
Sigma-Aldrich
苯酚, for molecular biology
Sigma-Aldrich
液状苯酚, ≥89.0%
Sigma-Aldrich
苯酚, BioXtra, ≥99.5% (GC)
Sigma-Aldrich
苯酚 溶液, Saturated with 0.1 M citrate buffer, pH 4.3 ± 0.2, BioReagent, for molecular biology
Sigma-Aldrich
苯酚, ≥99%
Supelco
苯酚, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
苯酚, contains hypophosphorous as stabilizer, loose crystals, ACS reagent, ≥99.0%
Sigma-Aldrich
苯酚, puriss., meets analytical specification of Ph. Eur., BP, USP, 99.5-100.5% (GC)
Sigma-Aldrich
苯酚, BioUltra, for molecular biology, TE-saturated, ~73% (T)
Sigma-Aldrich
苯酚, unstabilized, ReagentPlus®, ≥99%
Supelco
苯酚, PESTANAL®, analytical standard
Sigma-Aldrich
苯酚, puriss., meets analytical specification of Ph. Eur., BP, USP, ≥99.5% (GC), crystalline (detached)
Sigma-Aldrich
苯酚, ≥96.0% (calc. on dry substance, T)
Supelco
苯酚 溶液, 5000 μg/mL in methanol, certified reference material
Supelco
苯酚 溶液, certified reference material, 500 μg/mL in methanol
Sigma-Aldrich
苯酚, unstabilized, purified by redistillation, ≥99%
Sigma-Aldrich
苯酚, natural, 97%, FG
USP
苯酚, United States Pharmacopeia (USP) Reference Standard
Sigma-Aldrich
苯酚, BioUltra, for molecular biology, ≥99.5% (GC)
Supelco
苯酚 溶液, 100 μg/mL in acetonitrile, PESTANAL®, analytical standard
Sigma-Aldrich
抗 GLI1 (AB2) 兔抗, IgG fraction of antiserum
Sigma-Aldrich
Anti-Gli1 antibody produced in rabbit, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
Monoclonal Anti-GLI1 antibody produced in mouse, clone 3C8, purified immunoglobulin