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Epigenetics. 2017 Aug;12(8):698-707. doi: 10.1080/15592294.2017.1349045. Epub 2017 Jul 13.

Epigenetic silencing of miR-203 in Kazakh patients with esophageal squamous cell carcinoma by MassARRAY spectrometry.

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a Department of Pathology and Key Laboratory for Xinjiang Endemic and Ethnic Diseases , Shihezi University School of Medicine , Shihezi , China.
b Department of Pathology and Medical Research Center , Beijing Chaoyang Hospital, Capital Medical University , Beijing , China.
c Department of Pathology , Zhucheng Maternal and Child Care Service Centre , Weifang , China.
d Department of Pathology , People's Hospital of Wusu , Tacheng , China.
e Department of Thoracic and Cardiovascular Surgery , The First Affiliated Hospital, Shihezi University School of Medicine , Shihezi , China.
f Henan Key Laboratory for Esophageal Cancer Research , Department of Basic Oncology and Pathology at College of Medicine , The First and The Second Affiliated Hospital, Zhengzhou University , Zhengzhou , Henan , China.


Dysregulation of miR-203 by promoter methylation is associated with the development of various cancers. We aimed to explore the underlying link between promoter methylation and miR-203 expression in Kazakh esophageal squamous cell carcinoma (ESCC). MassARRAY® System spectrometry was used to quantitatively analyze the DNA methylation of 32 CpG sites within miR-203 in 99 Kazakh ESCC and 46 normal esophageal tissues (NETs) with similar population characteristics. We conducted real-time PCR to detect miR-203 expression levels and evaluated their association with methylation. Eleven CpG units within miR-203 promoter were frequently hypermethylated in ESCC compared with NETs (P < 0.05). The hypermethylation of several CpG units positively correlated with age, lower esophagus, constrictive type of ESCC, and moderately differentiated ESCC. Given the involvement of human papillomavirus (HPV) in etiology of ESCC was confirmed from our previous reports, herein we found that CpG units within miR-203 in HPV16-positive ESCC are more heavily methylated. Furthermore, miR-203 expression showed a nearly 4.5-fold decrease in ESCC than NETs (0.206 ± 0.336 vs. 0.908 ± 1.424, P < 0.001) and was significantly associated with lymph node metastasis (P = 0.012). The expression of miR-203 with 11 completely hypermethylated CpG units was approximately 6.5-fold lower than that with at least 1 unmethylated CpG unit (P < 0.001) and especially the CpG_15.16 and CpG_31.32 with higher methylation levels in ESCC tissues exhibited lower expression levels of miR-203, which indicated a reverse association between miR-203 methylation and expression. Hypermethylated miR-203 is a potential biomarker and targeted delivery of miR-203 could therefore serve as a preventive or therapeutic strategy for Kazakh ESCC.


Esophageal squamous cell carcinoma; Kazakh; methylation; miR-203

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