Zhiming Wang1, Shizhang Ling2, Eleni Rettig2, Ryan Sobel2, Marietta Tan2, Elana J Fertig3, Michael Considine3, Adel K El-Naggar4, Mariana Brait2, Carole Fakhry5, Patrick K Ha6. 1. Department of Oral and Maxillofacial Surgery, Shengjing Hospital of China Medical University, Shenyang, Liaoning, China; Department of Otolaryngology-Head and Neck Surgery, Johns Hopkins University, Baltimore, MD, USA. Electronic address: wangzm@sj-hospital.org. 2. Department of Otolaryngology-Head and Neck Surgery, Johns Hopkins University, Baltimore, MD, USA. 3. Department of Oncology Biostatistics, Johns Hopkins University, Baltimore, MD, USA. 4. Department of Pathology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. 5. Department of Otolaryngology-Head and Neck Surgery, Johns Hopkins University, Baltimore, MD, USA; Milton J. Dance Jr. Head and Neck Center at the Greater Baltimore Medical Center, Baltimore, MD, USA. 6. Department of Otolaryngology-Head and Neck Surgery, Johns Hopkins University, Baltimore, MD, USA; Milton J. Dance Jr. Head and Neck Center at the Greater Baltimore Medical Center, Baltimore, MD, USA. Electronic address: pha1@jhmi.edu.
Abstract
OBJECTIVES: The role of promoter methylation in the development of mucoepidermoid carcinoma (MEC) has not been fully explored. In this study, we investigated the epigenetic landscape of MEC. METHODS: The Illumina HumanMethylation27 BeadChip array and differential methylation analysis were utilized to screen for epigenetic alterations in 14 primary MEC tumors and 14 matched normal samples. Bisulfite sequencing was used to validate these results, with subsequent quantitative Methylation-Specific PCR (qMSP) to validate chloride intracellular channel protein 3 (CLIC3) in a separate cohort. Furthermore, CLIC3 immunohistochemical (IHC) staining was performed in another separate cohort of MEC. Finally, clinical and pathological characteristics were statistically analyzed for correlation with methylation status of CLIC3 and CLIC3 IHC H-scores by Wilcoxon rank sum, Kruskall-Wallis, and X(2) test tests. RESULTS: We obtained 6 significantly differentially methylated gene candidates demonstrating significant promoter hyper- or hypo-methylation from the array data. Using bisulfite sequencing, we found one gene, CLIC3, which showed differential methylation between MEC tumor and normal samples in a small validation cohort. qMSP analysis of the CLIC3 promoter in a separate validation set showed significantly lower methylation level in tumor than in normal. The level of CLIC3 methylation in MECs was not statistically correlated with clinical or pathological characteristics. However, IHC staining intensity and distribution of CLIC3 were significantly increased in MECs, compared with those of normal salivary gland tissues. CONCLUSIONS: Hypomethylation of CLIC3 promoter and its overexpression are significant events in MEC. Its functional role and potential therapeutic utility in MEC are worthy of further exploration.
OBJECTIVES: The role of promoter methylation in the development of mucoepidermoid carcinoma (MEC) has not been fully explored. In this study, we investigated the epigenetic landscape of MEC. METHODS: The Illumina HumanMethylation27 BeadChip array and differential methylation analysis were utilized to screen for epigenetic alterations in 14 primary MEC tumors and 14 matched normal samples. Bisulfite sequencing was used to validate these results, with subsequent quantitative Methylation-Specific PCR (qMSP) to validate chloride intracellular channel protein 3 (CLIC3) in a separate cohort. Furthermore, CLIC3 immunohistochemical (IHC) staining was performed in another separate cohort of MEC. Finally, clinical and pathological characteristics were statistically analyzed for correlation with methylation status of CLIC3 and CLIC3 IHC H-scores by Wilcoxon rank sum, Kruskall-Wallis, and X(2) test tests. RESULTS: We obtained 6 significantly differentially methylated gene candidates demonstrating significant promoter hyper- or hypo-methylation from the array data. Using bisulfite sequencing, we found one gene, CLIC3, which showed differential methylation between MEC tumor and normal samples in a small validation cohort. qMSP analysis of the CLIC3 promoter in a separate validation set showed significantly lower methylation level in tumor than in normal. The level of CLIC3 methylation in MECs was not statistically correlated with clinical or pathological characteristics. However, IHC staining intensity and distribution of CLIC3 were significantly increased in MECs, compared with those of normal salivary gland tissues. CONCLUSIONS: Hypomethylation of CLIC3 promoter and its overexpression are significant events in MEC. Its functional role and potential therapeutic utility in MEC are worthy of further exploration.
Authors: T T Money; R G King; M H Wong; J L Stevenson; B Kalionis; J J H M Erwich; M A Huisman; A Timmer; U Hiden; G Desoye; N M Gude Journal: Placenta Date: 2006-10-05 Impact factor: 3.481
Authors: Michelle D Williams; Nitin Chakravarti; Merrill S Kies; Shin-ichiro Maruya; Jeffrey N Myers; Joie C Haviland; Randal S Weber; Reuben Lotan; Adel K El-Naggar Journal: Clin Cancer Res Date: 2006-12-15 Impact factor: 12.531
Authors: Kowan Ja Jee; Marta Persson; Kristiina Heikinheimo; Fabricio Passador-Santos; Katri Aro; Sakari Knuutila; Edward W Odell; Antti Mäkitie; Kaarina Sundelin; Göran Stenman; Ilmo Leivo Journal: Mod Pathol Date: 2012-09-28 Impact factor: 7.842