TLR2 and TLR4 gene promoter methylation status during chronic periodontitis.
J Clin Periodontol
; 38(11): 975-83, 2011 Nov.
Article
in En
| MEDLINE
| ID: mdl-21899586
ABSTRACT
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 ANDMETHODS:
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.
Full text:
1
Database:
MEDLINE
Main subject:
Toll-Like Receptor 2
/
Toll-Like Receptor 4
/
Chronic Periodontitis
/
Gingiva
Type of study:
Observational_studies
Limits:
Adult
/
Female
/
Humans
/
Male
/
Middle aged
Language:
En
Year:
2011
Type:
Article