RRC ID 18663
著者 Kusumoto T, Hamada T, Yamada N, Nagata S, Kanmura Y, Houjou I, Kamikawa Y, Yonezawa S, Sugihara K.
タイトル Comprehensive epigenetic analysis using oral rinse samples: a pilot study.
ジャーナル J Oral Maxillofac Surg
Abstract PURPOSE:To prove that chromatin immunoprecipitation assay can be performed with oral rinse samples and to develop a protocol for comprehensive analysis of functional interactions among DNA methylation, histone modification, and gene expression using such samples.
MATERIALS AND METHODS:Eleven cancer cell lines and oral rinse samples from 10 patients with oral squamous cell carcinoma and 3 healthy subjects were examined. The expression of CDKN2A, a tumor suppressor gene, was determined by reverse transcription/polymerase chain reaction and immunohistochemistry. Promoter DNA methylation was assessed by methylation-specific polymerase chain reaction. Chromatin modifications were analyzed by a chromatin immunoprecipitation assay using antibodies for dimethylation and acetylation of lysine 9 of histone H3.
RESULTS:Epigenetic control of CDK2NA was observed in vitro in 11 cancer cell lines. Using the present protocol, comprehensive epigenetic analysis could be successfully performed with oral rinse samples. All patients were comfortable using the prescribed amount (16 mL) of normal saline to rinse their mouths. Nine patients (90%) and 1 healthy subject (33%) showed dimethylation of lysine 9 of histone H3. Moreover, 8 patients (80%) showed hypoacetylation of lysine 9 of histone H3, which was not observed in healthy subjects.
CONCLUSIONS:The present study showed for the first time that chromatin modifications can be analyzed using oral rinse samples by chromatin immunoprecipitation analysis. To evaluate the contribution of histone modifications for carcinogenesis of oral squamous cell carcinoma, studies including a larger number of subjects should be conducted in the future.
巻・号 70(6)
ページ 1486-94
公開日 2012-6-1
DOI 10.1016/j.joms.2011.04.021
PII S0278-2391(11)00775-0
PMID 21821326
MeSH Acetylation Adult Aged Aged, 80 and over Carcinoma, Squamous Cell / genetics* Carcinoma, Squamous Cell / metabolism Case-Control Studies Cell Line, Tumor Chromatin Immunoprecipitation / methods* Cyclin-Dependent Kinase Inhibitor p16 / biosynthesis Cyclin-Dependent Kinase Inhibitor p16 / genetics* DNA Methylation Epigenesis, Genetic* Female Gene Expression Regulation, Neoplastic Gene Silencing Genes, p16 / physiology Histones / genetics* Histones / metabolism Humans Lysine / genetics Lysine / metabolism Male Middle Aged Mouth Mucosa / pathology* Mouth Neoplasms / genetics* Mouth Neoplasms / metabolism Pilot Projects Promoter Regions, Genetic / genetics
IF 1.642
引用数 9
WOS 分野 DENTISTRY, ORAL SURGERY & MEDICINE
リソース情報
ヒト・動物細胞 HSC-3(RCB1975) HSC-4(RCB1902) Ca9-22(RCB1976)