Comprehensive Epigenetic Analysis Using Oral Rinse Samples: A Pilot Study
| Autor: | Yuji Kanmura, Satoshi Nagata, Norishige Yamada, Yoshiaki Kamikawa, Kazumasa Sugihara, Suguru Yonezawa, Takanobu Kusumoto, Tomofumi Hamada, Izumi Houjou |
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| Rok vydání: | 2012 |
| Předmět: |
Adult
Male Chromatin Immunoprecipitation Pilot Projects medicine.disease_cause Epigenesis Genetic Histones Histone H3 Cell Line Tumor medicine Humans Gene Silencing Epigenetics Promoter Regions Genetic Cyclin-Dependent Kinase Inhibitor p16 Aged Aged 80 and over biology business.industry Genes p16 Lysine Mouth Mucosa Acetylation DNA Methylation Middle Aged Molecular biology Chromatin Gene Expression Regulation Neoplastic Histone Otorhinolaryngology Case-Control Studies DNA methylation Carcinoma Squamous Cell biology.protein Female Mouth Neoplasms Surgery Oral Surgery Carcinogenesis business Chromatin immunoprecipitation |
| Zdroj: | Journal of Oral and Maxillofacial Surgery. 70:1486-1494 |
| ISSN: | 0278-2391 |
| DOI: | 10.1016/j.joms.2011.04.021 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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