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Gli Regulates MUC5AC Transcription in Human Gastrointestinal Cells

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...

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Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4148389/
https://www.ncbi.nlm.nih.gov/pubmed/25166306
http://dx.doi.org/10.1371/journal.pone.0106106
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author 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
author_facet 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
author_sort Kageyama-Yahara, Natsuko
collection PubMed
description 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.
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spelling pubmed-41483892014-08-29 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 PLoS One Research Article 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. Public Library of Science 2014-08-28 /pmc/articles/PMC4148389/ /pubmed/25166306 http://dx.doi.org/10.1371/journal.pone.0106106 Text en © 2014 Kageyama-Yahara et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
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
title Gli Regulates MUC5AC Transcription in Human Gastrointestinal Cells
title_full Gli Regulates MUC5AC Transcription in Human Gastrointestinal Cells
title_fullStr Gli Regulates MUC5AC Transcription in Human Gastrointestinal Cells
title_full_unstemmed Gli Regulates MUC5AC Transcription in Human Gastrointestinal Cells
title_short Gli Regulates MUC5AC Transcription in Human Gastrointestinal Cells
title_sort gli regulates muc5ac transcription in human gastrointestinal cells
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4148389/
https://www.ncbi.nlm.nih.gov/pubmed/25166306
http://dx.doi.org/10.1371/journal.pone.0106106
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