Gli regulates MUC5AC transcription in human gastrointestinal cells

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

Research output: Contribution to journalArticle

10 Citations (Scopus)

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 Glibinding 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.

Original languageEnglish
Article numbere106106
JournalPLoS One
Volume9
Issue number8
DOIs
Publication statusPublished - 28-08-2014

Fingerprint

Transcription
transcription (genetics)
Genes
Cells
promoter regions
Luciferases
Genetic Promoter Regions
Gene expression
Assays
Cell Line
Tissue
cell lines
luciferase
Histone Deacetylase Inhibitors
Mammals
5' Flanking Region
DNA
Differentiation Antigens
cells
genes

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Kageyama-Yahara, N., Yamamichi, N., Takahashi, Y., Nakayama, C., Shiogama, K., Inada, K. I., ... Koike, K. (2014). Gli regulates MUC5AC transcription in human gastrointestinal cells. PLoS One, 9(8), [e106106]. https://doi.org/10.1371/journal.pone.0106106
Kageyama-Yahara, Natsuko ; Yamamichi, Nobutake ; Takahashi, Yu ; Nakayama, Chiemi ; Shiogama, Kazuya ; Inada, Ken Ichi ; Konno-Shimizu, Maki ; Kodashima, Shinya ; Fujishiro, Mitsuhiro ; Tsutsumi, Yutaka ; Ichinose, Masao ; Koike, Kazuhiko. / Gli regulates MUC5AC transcription in human gastrointestinal cells. In: PLoS One. 2014 ; Vol. 9, No. 8.
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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 Glibinding 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.",
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Kageyama-Yahara, N, Yamamichi, N, Takahashi, Y, Nakayama, C, Shiogama, K, Inada, KI, Konno-Shimizu, M, Kodashima, S, Fujishiro, M, Tsutsumi, Y, Ichinose, M & Koike, K 2014, 'Gli regulates MUC5AC transcription in human gastrointestinal cells', PLoS One, vol. 9, no. 8, e106106. https://doi.org/10.1371/journal.pone.0106106

Gli regulates MUC5AC transcription in human gastrointestinal cells. / Kageyama-Yahara, Natsuko; Yamamichi, Nobutake; Takahashi, Yu; Nakayama, Chiemi; Shiogama, Kazuya; Inada, Ken Ichi; Konno-Shimizu, Maki; Kodashima, Shinya; Fujishiro, Mitsuhiro; Tsutsumi, Yutaka; Ichinose, Masao; Koike, Kazuhiko.

In: PLoS One, Vol. 9, No. 8, e106106, 28.08.2014.

Research output: Contribution to journalArticle

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T1 - Gli regulates MUC5AC transcription in human gastrointestinal cells

AU - Kageyama-Yahara, Natsuko

AU - Yamamichi, Nobutake

AU - Takahashi, Yu

AU - Nakayama, Chiemi

AU - Shiogama, Kazuya

AU - Inada, Ken Ichi

AU - Konno-Shimizu, Maki

AU - Kodashima, Shinya

AU - Fujishiro, Mitsuhiro

AU - Tsutsumi, Yutaka

AU - Ichinose, Masao

AU - Koike, Kazuhiko

PY - 2014/8/28

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N2 - 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 Glibinding 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.

AB - 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 Glibinding 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.

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Kageyama-Yahara N, Yamamichi N, Takahashi Y, Nakayama C, Shiogama K, Inada KI et al. Gli regulates MUC5AC transcription in human gastrointestinal cells. PLoS One. 2014 Aug 28;9(8). e106106. https://doi.org/10.1371/journal.pone.0106106