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DNA methylation status of the IL8 gene promoter in oral cells of smokers and non‐smokers with chronic periodontitis
Author(s) -
Oliveira Naila F. P.,
Damm Gilcy R.,
Andia Denise C.,
Salmon Cristiane,
Nociti Jr. Francisco H.,
Line Sérgio R. P.,
De Souza Ana Paula
Publication year - 2009
Publication title -
journal of clinical periodontology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.456
H-Index - 151
eISSN - 1600-051X
pISSN - 0303-6979
DOI - 10.1111/j.1600-051x.2009.01446.x
Subject(s) - interleukin 8 , dna methylation , chronic periodontitis , periodontitis , methylation , polymerase chain reaction , microbiology and biotechnology , oral mucosa , inflammation , bisulfite sequencing , interleukin , genomic dna , gene , real time polymerase chain reaction , medicine , biology , gene expression , immunology , pathology , cytokine , genetics
Aim: This study analysed the status of DNA methylation in the promoter region of the IL8 gene in oral mucosa cells from healthy, smoker and non‐smoker subjects with chronic periodontitis and compared these findings among groups with mRNA levels. Material and Methods: Genomic DNA from epithelial oral cells of 41 healthy subjects, 30 smokers with chronic periodontitis and 40 non‐smokers with chronic periodontitis were purified and modified by sodium bisulphite. Genomic DNA from blood leucocytes and gingival cells from biopsies of 13 subjects of each group were also purified and modified by sodium bisulphite. Modified DNA was submitted by methylation‐specific polymerase chain reaction (PCR) (MSP), electrophoresed on 10% polyacrylamide gels and stained with SYBR Gold. Total RNA from gingival cells was also isolated using the TRIzol reagent, and real‐time PCR performance was used to detect the levels of interleukin‐8 mRNA. Results: Our results indicate that individuals with chronic periodontitis, independent of smoking habit, have a higher percentage of hipomethylation of the IL8 gene than those controls in epithelial oral cells ( p <0.0001), and expression of higher levels of interleukin‐8 (IL‐8) mRNA than controls in gingival cells ( p= 0.007). No significant differences among groups were observed in gingival cells and blood cells. Conclusion: We conclude that inflammation in the oral mucosa might lead to changes in the DNA methylation status of the IL8 gene in epithelial oral cells.