title: Clinical significance of epigenetic silencing and re-expression of O6-methylguanine-DNA methyltransferase using epigenetic agents in laryngeal carcinoma.
abstract: The aim of the present study was to investigate the association between O6-methylguanine-DNA methyltransferase (MGMT) gene expression levels, and DNA methylation status and histone modifications in laryngeal squamous cell carcinoma (LSCC). Chromatin immunoprecipitation, methylation-specific polymerase chain reaction (PCR), and reverse transcription-quantitative PCR were performed to analyze histone modifications, DNA methylation status and mRNA expression levels in the promoter region of the MGMT gene in laryngeal carcinoma HEp-2 cells, as well as in 50 paired healthy and LSCC tissue samples. The present study demonstrated that treatment of HEp-2 cells with 5-aza-2'-deoxycytidine (Aza), a DNA methyltransferase inhibitor, significantly upregulated MGMT mRNA expression levels, reduced MGMT DNA methylation, reduced MGMT histone H3 lysine 9 (H3K9) di-methylation, and increased MGMT histone H3 lysine 4 di-methylation without a significant change in H3K9 acetylation. Trichostatin A (TSA), a histone deacetylase inhibitor, marginally upregulated MGMT mRNA expression levels without affecting the DNA methylation status, or H3K9 or H3K4 di-methylation, however, TSA treatment caused a significant increase in H3K9 acetylation. Furthermore, Aza and TSA combination treatment produced a synergistic effect. In the LSCC samples, the rate of DNA methylation in the MGMT gene was 54%, compared with 24% in the healthy control group (P<0.05). Therefore, data from the present study indicates that MGMT may serve as a novel therapeutic target in the treatment of LSCC.
authors.affiliation: Department of Otorhinolaryngology, Shengjing Hospital of China Medical University, Shenyang, Liaoning 110004, P.R. China.
authors.forename: Jing
authors.initials: J
authors.lastname: Yang
authors.affiliation: Department of General Sugery, General Hospital of Liaohe Oil Field, Panjin, Liaoning 124010, P.R. China.
authors.forename: Xin-Bing
authors.initials: XB
authors.lastname: Zhu
authors.affiliation: Department of Otorhinolaryngology, Fushun Second Hospital, Fushun, Liaoning 113001, P.R. China.
authors.forename: Li-Xia
authors.initials: LX
authors.lastname: He
authors.affiliation: Department of Otorhinolaryngology, Shengjing Hospital of China Medical University, Shenyang, Liaoning 110004, P.R. China.
authors.forename: Zhao-Wei
authors.initials: ZW
authors.lastname: Gu
authors.affiliation: Department of Otorhinolaryngology, Shengjing Hospital of China Medical University, Shenyang, Liaoning 110004, P.R. China.
authors.forename: Ming-Zhu
authors.initials: MZ
authors.lastname: Jin
authors.affiliation: Department of Otorhinolaryngology, Shengjing Hospital of China Medical University, Shenyang, Liaoning 110004, P.R. China.
authors.forename: Wen-Yue
authors.initials: WY
authors.lastname: Ji
keywords: 5-aza-2′-deoxycytidine; DNA methylation; O6-methylguanine-DNA methyltransferase gene; histone modification; laryngeal carcinoma; trichostatin A