Epigenetic alterations of the keratin 13 gene in oral squamous cell carcinoma

BMC Cancer

Kaori Naganuma¹  Mitsutoki Hatta²  Tetsuro Ikebe¹  Jun Yamazaki² 

[1] Department of Oral and Maxillofacial Surgery, Fukuoka Dental College, Fukuoka 814-0193, Japan

[2] Department of Physiological Science and Molecular Biology, Fukuoka Dental College, Fukuoka 814-0193, Japan

关键词: Gene silencing;    Polycomb repressive complex 2 (PRC2);    Oral squamous cell carcinoma (OSCC);    Keratin 13 (KRT13);   

Others  :  1106809
DOI  :  10.1186/1471-2407-14-988

 received in 2014-08-08, accepted in 2014-12-16,  发布年份 2014

【 摘 要 】

Background

Epigenetic modifications play important roles in the regulation of gene expression determining cellular phenotype as well as various pathologies such as cancer. Although the loss of keratin 13 (KRT13) is reportedly linked to malignant transformation of oral epithelial cells, the molecular mechanisms through which KRT13 is repressed in oral squamous cell carcinoma (OSCC) remain unclear. The aim of this study is to identify the epigenetic alterations of the KRT13 gene in OSCCs.

Methods

We investigated KRT13 expression levels and chromatin modifications of the KRT13 promoter in the three OSCC cell lines (HSC4, HSC3, and SAS). The expression levels of KRT13 protein and mRNA were analyzed by western blotting and quantitative reverse-transcription polymerase chain reaction, respectively, and the localization of KRT13 protein was detected by immunofluorescence. DNA methylation and histone modifications in the KRT13 promoter were determined by bisulfite sequencing and chromatin immunoprecipitation (ChIP), respectively. For the pharmacological depletion of Polycomb repressive complex 2 (PRC2), cells were treated with 3-deazaneplanocin A (DZNep).

Results

KRT13 expression was transcriptionally silenced in the HSC3 and SAS cells and post-transcriptionally repressed in the HSC4 cells, while the KRT13 promoter was hypermethylated in all of the three OSCC cell lines. ChIP analysis revealed that PRC2-mediated trimethylation of Lys 27 on histone H3 (H3K27me3) was increased in the KRT13 promoter in the HSC3 and SAS cells. Finally, we demonstrated that the treatment of SAS cells with DZNep reactivated the transcription of KRT13 gene.

Conclusions

Our data provide mechanistic insights into the epigenetic silencing of KRT13 genes in OSCC cells and might be useful for the development of diagnostic markers and novel therapeutic approaches against OSCCs.

【 授权许可】

   
2014 Naganuma et al.; licensee BioMed Central.

【 图 表 】

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