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Transient receptor potential vanilloid 4 ( TRPV 4) silencing in Helicobacter pylori‐ infected human gastric epithelium
Author(s) -
Mihara Hiroshi,
Suzuki Nobuhiro,
Muhammad Jibran Sualeh,
Nanjo Sohachi,
Ando Takayuki,
Fujinami Haruka,
Kajiura Shinya,
Hosokawa Ayumu,
Sugiyama Toshiro
Publication year - 2017
Publication title -
helicobacter
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.206
H-Index - 79
eISSN - 1523-5378
pISSN - 1083-4389
DOI - 10.1111/hel.12361
Subject(s) - gene silencing , methylation , dna methylation , microbiology and biotechnology , helicobacter pylori , epithelium , biology , gastric mucosa , chemistry , gene expression , stomach , biochemistry , gene , genetics
Background Helicobacter pylori ( HP ) infection induces methylation silencing of specific genes in gastric epithelium. Various stimuli activate the nonselective cation channel TRPV 4, which is expressed in gastric epithelium where it detects mechanical stimuli and promotes ATP release. As CpG islands in TRPV 4 are methylated in HP ‐infected gastric epithelium, we evaluated HP infection‐dependent changes in TRPV 4 expression in gastric epithelium. Materials and Methods Human gastric biopsy samples, a human gastric cancer cell line ( AGS ), and a normal gastric epithelial cell line ( GES ‐1) were used to detect TRPV 4 mRNA and protein expression by RT ‐ PCR and Western blotting, respectively. Ca 2+ imaging was used to evaluate TRPV 4 ion channel activity. TRPV 4 methylation status was assessed by methylation‐specific PCR ( MSP ). ATP release was measured by a luciferin‐luciferase assay. Results TRPV 4 mRNA and protein were detected in human gastric biopsy samples and in GES ‐1 cells. MSP and demethylation assays showed TRPV 4 methylation silencing in AGS cells. HP coculture directly induced methylation silencing of TRPV 4 in GES ‐1 cells. In human samples, HP infection was associated with TRPV 4 methylation silencing that recovered after HP eradication in a time‐dependent manner. Conclusion HP infection‐dependent DNA methylation suppressed TRPV 4 expression in human gastric epithelia, suggesting that TRPV 4 methylation may be involved in HP ‐associated dyspepsia.