mRNA expression and hypermethylation of tumor suppressor genes apoptosis protease activating factor-1 and death-associated protein kinase in oral squamous cell carcinoma

Li,Chunyan; Wang,Lin; Su,Jing; Zhang,Ruhui; Fu,Li; Zhou,Yanmin

doi:10.3892/ol.2013.1353

mRNA expression and hypermethylation of tumor suppressor genes apoptosis protease activating factor-1 and death-associated protein kinase in oral squamous cell carcinoma

Authors:
- Chunyan Li
- Lin Wang
- Jing Su
- Ruhui Zhang
- Li Fu
- Yanmin Zhou
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Affiliations: Department of Implant Center, School of Dentistry, Jilin University, Changchun, Jilin 130011, P.R. China, Department of VIP Integration, School of Dentistry, Jilin University, Changchun, Jilin 130011, P.R. China, Department of Pathophysiology, School of Basic Medical Sciences, Jilin University, Changchun, Jilin 130011, P.R. China, Department of Oral and Maxillofacial Surgery, School of Dentistry, Jilin University, Changchun, Jilin 130011, P.R. China
Published online on: May 17, 2013 https://doi.org/10.3892/ol.2013.1353
Pages: 280-286

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Abstract

Apoptosis protease activating factor‑1 (Apaf‑1) and death‑associated protein kinase (DAPK) are p53 pathway‑related genes that play significant roles in the activation of caspases, which are involved in mitochondrial‑mediated apoptosis. The present study aimed to confirm the role of hypermethylation of the Apaf‑1 and DAPK gene promoter regions in oral squamous cell carcinoma (OSCC) and the effect of the demethylation drug, 5‑aza‑2'‑deoxycytidine (DAC). mRNA from 53 OSCC samples, 23 normal oral mucosa samples and Tca8113 human tongue carcinoma cell lines was detected using semi‑quantitative reverse transcription‑polymerase chain reaction (RT‑PCR). The DNA from each sample was analyzed using methylation‑specific PCR (MSP). The Tca8113 cells were demethylated using DAC and the demethylation and re‑expression of Apaf‑1 and DAPK were analyzed. The Apaf‑1 and DAPK mRNA expression index was decreased in 51 (96.23%) and 50 (94.34%) cases, respectively, in the tumor tissues. Hypermethylation of the Apaf‑1 and DAPK promoter regions was detected in 46 (86.79%) and 38 (71.69%) cases, respectively. Promoter hypermethylation of the two genes correlated with a decreased mRNA expression in the tumor tissues. Subsequent to being treated with DAC, Apaf‑1 and DAPK were demethylated and re‑expressed in the Tca8113 cells. Apaf‑1 and DAPK promoter hypermethylation may be associated with low gene expression in OSCC. Furthermore, a loss of Apaf‑1 and DAPK expression may recover following demethylation. The data provide evidence that methylation exists in OSCC and may play a role in the development of this disease.

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