DNA methylation status of the IL8 gene promoter in oral cells of smokers and non-smokers with chronic periodontitis
DNA methylation status of the IL8 gene promoter in oral cells of smokers and non-smokers with chronic periodontitis
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DOI：
10.1111/j.1600-051x.2009.01446.x
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发表时间：
2009-09-01
影响因子：
6.7
通讯作者：
de Souza, Ana Paula
中科院分区：
文献类型：
--
作者：
Oliveira, Naila F. P.;Damm, Gilcy R.;de Souza, Ana Paula
目的分析慢性牙周炎患者、吸烟者和非吸烟者口腔黏膜细胞中IL8基因启动子区DNA甲基化状态，并进行组间比较。材料与方法用亚硫酸氢钠对41例健康人、30例吸烟者和40例非吸烟者口腔粘膜上皮细胞基因组DNA进行纯化和修饰。取每组13例受试者的外周血白细胞和牙周组织，提取基因组DNA，用亚硫酸氢钠进行修饰。用甲基化特异性聚合酶链式反应(MSP)将DNA修饰后，在10%的聚丙烯酰胺凝胶上进行凝胶电泳，SYBR Gold染色。结果与吸烟习惯无关的慢性牙周炎患者口腔上皮细胞IL-8基因甲基化百分率高于对照组(p<；0.0001)，IL-8基因表达水平高于对照组(p=0.007)。结论口腔黏膜炎症可能导致口腔上皮细胞IL8基因甲基化状态的改变。
AimThis 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 MethodsGenomic 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.ResultsOur 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.ConclusionWe conclude that inflammation in the oral mucosa might lead to changes in the DNA methylation status of the IL8 gene in epithelial oral cells.