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Expression and methylation status of microRNA-203 gene in tissues and cells of esophageal squamous cell carcinoma
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Abstract:
Objective: To investigate the expression, methylation status and functional role of miR-203 in pathogenesis of ESCC. ethods: Eighty-three patients diagnosed with ESCC in Hebei Medical University-Affiliated Fourth Hospital between 2008 and 2011 were recruited. Biopsy specimens were collected from primary tumors and the corresponding adjacent tissues. Quantitative real-time RT-PCR (qRT-PCP) and methylation specific PCR (MSP) were used to respectively detect the mRNA abundance and methylation status of miR-203 gene in the collected specimens. Five esophageal cancer cell lines (TE1, TE13, T.TN, Yes-2, and EC109) were treated with DNA methyltransferase inhibitor 5-Aza-2’-detoxycytidine (5-Aza-dC) Levels of CpG methylation of the miR-203 gene and miR-203 were assessed by qRT-PCR and MSP, respectively, 72 h after 5-Aza-dC treatment. Results： Relatively low levels of miR-203 mRNA and hypermethylation were detected in all the five untreated esophageal cancer cell lines. After 5-Aza-dC treatment, miR-203 mRNA was increased in all five cell lines studied and the methylation level of miR-203 was decreased in YES-2 cells and complete miR-203 unmethylation occurred in TE1, TE13, T.TN, and EC109 cells. The abundance of miR-203 mRNA was significantly lower (0.54±0.11 vs 1.00±0.01, P<0.05) and the methylation frequency of miR-203 promoter was significantly higher (62.65%vs 7.23%,P<0.05) in ESCC tissues than in corresponding tissues. Both miR-203 mRNA abundance and methylation frequency were all correlated with TNM stage and pathological differentiation (P<0.05). The expression of miR-203 in ESCC with miR-203 methylation was significantly lower than that in ESCC without miR-203 methylation (P<0.05). Conclusion： Aberrantly low expression of miR-203 is closely related to the development and progression of ESCC and promoter DNA methylation is one of the possible mechanisms underlying miR-203 inactivationin ESCC.
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Project Supported:
Project supported by the National Natural Science Foundation（No. 81101854）, and the Major Projects of Medical Research Foundation of Hebei Province （No. \[2012\]2056）
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