TLR2 and TLR4 gene promoter methylation status during chronic periodontitis

Autor(es)

Paulo De Oliveira, Naila Francis ; Andia, Denise Carleto ; Planello, Aline Cristiane ; Pasetto, Silvana ; Marques, Marcelo Rocha ; Nociti, Francisco Humberto, Jr. ; Peres Line, Sergio Roberto ; De Souza, Ana Paula

• Tipo de documento: Artigo Científico
• Fonte: JOURNAL OF CLINICAL PERIODONTOLOGY; v. 38, n. 11, p. 9-pg., 2011-11-01.

Resumo

Aim: The objective of this study was to analyse the status of DNA methylation in the promoter region of the toll-like receptor (TLR) 2 and TLR4 genes in gingival tissue samples from healthy subjects, smokers and non-smokers affected by chronic periodontitis. Material and Methods: Genomic DNA and total RNA were purified from gingival tissue using the TRIZOL reagent protocol. Genomic DNA was then digested by methylation-sensitive restriction enzymes, amplified by polymerase chain reaction (PCR), electrophoresed on a 10% polyacrylamide gel and stained using SYBR Gold. Real-time PCR was also performed to verify the transcript levels. Results: The CpG dinucleotides analysed were observed to be unmethylated in the majority of DNA samples of the three groups and statistical differences were not found among groups (p > 0.05). However, a trend towards methylation was observed in the TLR2 HhaI site in the samples of the periodontitis non-smoker groups. In fact, the analysis of all CpG sites together shows which complete methylation is observed in the shortest level in the samples of periodontitis non-smoker group. The analysis of transcript levels demonstrated no difference among groups (p > 0.05). Conclusion: The results demonstrated major unmethylation of the TLR4 gene promoter in all groups. However, the results for the TLR2 gene promoter are inconclusive; this gene was found as a mosaic of methylated and unmethylated DNA in the majority of samples of the three groups and we also observed a trend towards the DNA methylation of CpG sites recognized by the HhaI enzyme. (AU)

• Processo FAPESP: 07/02488-0 - Análise de metilação no promotor dos genes da IL-8, iNOS, TIMP-2 e TIMP-3 em indivíduos fumantes e não-fumantes com periodontite crônica: associação com a expressão gênica destes genes no tecido gengival
• Beneficiário: Ana Paula de Souza
• Modalidade de apoio: Auxílio à Pesquisa - Regular