title: DNA hypermethylation of extracellular matrix-related genes in human periodontal fibroblasts induced by stimulation for a prolonged period with lipopolysaccharide derived from Porphyromonas gingivalis.
abstract: OBJECTIVE
  The involvement of DNA methylation in periodontal disease is not clear. Lipopolysaccharide (LPS) derived from Porphyromonas gingivalis is involved in the progression of periodontal disease. We recently developed an in vitro model of LPS infection in human periodontal fibroblast cells (HPdLFs) for a prolonged period. In this study, we examined genome-wide analysis of DNA methylation in HPdLFs stimulated with LPS derived from P. gingivalis for a prolonged period. We noted the hypermethylation of extracellular matrix (ECM)-related genes and examined whether hypermethylation affected their transcription levels.
  
  
  METHODS
  HPdLFs were grown in Dulbecco's modified Eagle's medium containing 10% fetal bovine serum. The culture was repeated, alternating 3 d with LPS derived from P. gingivalis and 3 d without LPS for 1 mo. Untreated samples were used as controls. DNA was analyzed using the human CpG island microarray. Quantitative methylation-specific polymerase chain reaction was carried out to confirm reproducibility of the microarray data. The expression levels of mRNA of the selected ECM-related genes from the data were analyzed by quantitative reverse transcription-polymerase chain reaction.
  
  
  RESULTS
  We found 25 ECM-related genes with hypermethylation at the CpG island of the promoter region, which exhibited a fourfold greater hypermethylation than controls. Among these genes, hypermethylation of nine ECM-related genes, FANK1, COL4A1-A2, 12A1 and 15A1, LAMA5 and B1, MMP25, POMT1 and EMILIN3, induced a significantly downregulated expression of their mRNA.
  
  
  CONCLUSIONS
  These results indicate that LPS derived from P. gingivalis may cause DNA hypermethylation of some ECM-related genes followed by downregulated expression of their transcriptional levels.
authors.affiliation: Division of Oral Medicine and Pathology, Department of Human Biology and Pathophysiology, Health Sciences University of Hokkaido, Ishikari-Tobetsu, Hokkaido, Japan.
authors.forename: R
authors.initials: R
authors.lastname: Takai
authors.affiliation: Division of Disease Control and Molecular Epidemiology, Department of Oral Growth and Development, Health Sciences University of Hokkaido, Ishikari-Tobetsu, Hokkaido, Japan.
authors.forename: O
authors.initials: O
authors.lastname: Uehara
authors.affiliation: Division of Oral Medicine and Pathology, Department of Human Biology and Pathophysiology, Health Sciences University of Hokkaido, Ishikari-Tobetsu, Hokkaido, Japan.
authors.forename: F
authors.initials: F
authors.lastname: Harada
authors.affiliation: Division of Oral Medicine and Pathology, Department of Human Biology and Pathophysiology, Health Sciences University of Hokkaido, Ishikari-Tobetsu, Hokkaido, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Utsunomiya
authors.affiliation: Division of Oral Medicine and Pathology, Department of Human Biology and Pathophysiology, Health Sciences University of Hokkaido, Ishikari-Tobetsu, Hokkaido, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Chujo
authors.affiliation: Division of Oral Medicine and Pathology, Department of Human Biology and Pathophysiology, Health Sciences University of Hokkaido, Ishikari-Tobetsu, Hokkaido, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Yoshida
authors.affiliation: Division of Oral Medicine and Pathology, Department of Human Biology and Pathophysiology, Health Sciences University of Hokkaido, Ishikari-Tobetsu, Hokkaido, Japan.
authors.forename: J
authors.initials: J
authors.lastname: Sato
authors.affiliation: Division of Oral Medicine and Pathology, Department of Human Biology and Pathophysiology, Health Sciences University of Hokkaido, Ishikari-Tobetsu, Hokkaido, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Nishimura
authors.affiliation: Division of Disease Control and Molecular Epidemiology, Department of Oral Growth and Development, Health Sciences University of Hokkaido, Ishikari-Tobetsu, Hokkaido, Japan.
authors.forename: I
authors.initials: I
authors.lastname: Chiba
authors.affiliation: Division of Oral Medicine and Pathology, Department of Human Biology and Pathophysiology, Health Sciences University of Hokkaido, Ishikari-Tobetsu, Hokkaido, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Abiko
mesh_terms: D002478:Cells, Cultured; D019175:DNA Methylation; D015536:Down-Regulation; D005109:Extracellular Matrix / Q000235:genetics* / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism*; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology*; D016966:Porphyromonas gingivalis; D014158:Transcription, Genetic
chemical_list: D008070:Lipopolysaccharides
keywords: DNA methylation; extracellular matrix; lipopolysaccharide; periodontal fibroblasts