Surgical oncology and reconstructionComprehensive Epigenetic Analysis Using Oral Rinse Samples: A Pilot Study
Section snippets
Cell Lines
Human tongue SCC cell lines HSC-3 and HSC-4 and human gingival carcinoma cell line Ca9-22 were obtained from RIKEN BioResource Center (Ibaraki, Japan). Human breast cancer (MCF-7 and T-47D), human lung cancer (A427 and NCI-H292), human pancreatic carcinoma (BxPC3 and HPAFII), and human colon adenocarcinoma (LS174T and Caco2) cell lines were obtained from American Type Culture Collection (Manassas, VA). HSC-3, Ca9-22, MCF-7, A427, HPAFII, LS174T, and Caco2 cells were cultured in Eagle's Minimum
Expression and Epigenetic Status of CDKN2A in Cancer Cell Lines
To confirm the previously reported epigenetic mechanism of CDKN2A and establish an experimental method for further study using oral rinse samples, RT-PCR, MSP, and ChIP analyses were performed using 11 human cancer cell lines. HSC-3, HSC-4, and HPAFII cells expressed CDKN2A mRNA, as examined by RT-PCR analysis (Fig 1A), but LS174T cells showed relatively low expression. Ca9-22, MCF-7, T-47D, A427, NCI-H292, BxPC-3, and Caco2 cells did not show CDKN2A mRNA expression.
The status of DNA promoter
Discussion
DNA methylation and chromatin modification are essential for regulating gene activity.8 DNA methylation effectively downregulates gene activity by the addition of a methyl group to the 5-carbon of a cytosine base. Less specifically, modification of the chromatin structure can be achieved by multiple mechanisms that cause upregulation or downregulation of the associated gene.3, 21 Pathologic events such as carcinogenesis can be caused by a drastic alteration in established methylation patterns
Acknowledgments
This work was supported by Grants-in-Aid 19390521 (K. Sugihara), 19791533 (T. Hamada), 21792020 (T. Hamada) and 20014022 (S. Yonezawa) from the Ministry of Education, Culture, Sports, Science and Technology of Japan and by the JSPS Fellowship Grant-in-Aid 219447 (N. Yamada).
References (29)
Global epidemiology of oral and oropharyngeal cancer
Oral Oncol
(2009)The essentials of DNA methylation
Cell
(1992)- et al.
Inactivation of the p14(ARF), p15(INK4B) and p16(INK4A) genes is a frequent event in human oral squamous cell carcinomas
Oral Oncol
(2001) - et al.
Gene promoter hypermethylation in oral rinses of leukoplakia patients—A diagnostic and/or prognostic tool?
Eur J Cancer
(2003) Saliva as a tool for oral cancer diagnosis and prognosis
Oral Oncol
(2009)- et al.
p16(MTS-1/CDKN2/INK4a) in cancer progression
Exp Cell Res
(2001) - et al.
Inactivation patterns of the p16 (INK4a) gene in oral squamous cell carcinoma cell lines
Oral Oncol
(1999) - et al.
Global cancer statistics, 2002
CA Cancer J Clin
(2005) - et al.
MUC2 expression is regulated by histone H3 modification and DNA methylation in pancreatic cancer
Int J Cancer
(2006) - et al.
MUC1 expression is regulated by DNA methylation and histone H3 lysine 9 modification in cancer cells
Cancer Res
(2008)
Triple analysis of the cancer epigenome: An integrated microarray system for assessing gene expression, DNA methylation, and histone acetylation
Cancer Res
Silencing of transgene transcription precedes methylation of promoter DNA and histone H3 lysine 9
EMBO J
An overview of epigenetic assays
Mol Biotechnol
The state-of-the-art of chromatin immunoprecipitation
Methods Mol Biol
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2018, Journal of Oral BiosciencesCitation Excerpt :In contrast to that acetylation of H3 histone was also observed at specific lysine molecule positions (k4 = 37%, k9 = 80%, k18 = 39% of oral cancers samples analyzed). The percentages/ratios for lysine molecule positions were calculated by dividing the total number of methylated samples with the total number of samples being analyzed [40–43]. Furthermore, the correlations were drawn between histone modifications and clinicopathological parameters like nodal status, the size of the tumor, tumor stage, perineural invasion in oral squamous cell carcinoma.
Clinical implications of epigenetic regulation in oral cancer
2015, Oral OncologyCitation Excerpt :Cumulative studies on histone modifications in oral cancers demonstrated methylation of H3 histone at specific lysine molecule positions- K4 in 39% (n = 145), K9 in 47% (n = 199) and K27 in 45% (n = 160) oral cancers. Acetylation of lysines on H3 histone was reported at positions K4 in 37% (n = 171), K9 in 80% (n = 10) and K18 in 39% (n = 152) of oral cancers [98–100]. Besides, methylation at H3K27 correlated with the clinico-pathological parameters i.e. size of tumor, nodal status, tumor stage and perineural invasion [99].
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Role of epigenetics in OSCC: an understanding above genetics
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