Expression and methylation status of miR-6867 and its host gene RAPGEFL1 in human esophageal squamous cell carcinoma tissues and cell lines
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Abstract:
Objective:To detect the expression and methylation status of miR-6867 and its host gene RAPGEFL1 (rap guanine nucleotide exchange factor like 1) in human esophageal carcinoma cell lines and esophageal squamous cell carcinoma (ESCC) tissues, and to explore the role of miR-6867 and RAPGEFL1 in occurrence and development of ESCC. Methods: :Tissue specimens were obtained from 87 ESCC patients treated in the Fourth Affiliated Hospi-tal of Hebei Medical University between January 2014 and January 2016. The expression of miR-6867 and RAPGE-FL1 gene in esophageal cancer cell lines and ESCC tissues as well as the corresponding noncancerous tissues were detected by real-time fluorescent quantitative PCR. The relationship of miR-6867 and RAPGEFL1 gene expression were analyzed. The methylation status of RAPGEFL1 in esophageal cancer cell lines and ESCC tissues and the cor-responding noncancerous tissues was detected by methylation specific PCR methods. Results: :The relative expres-sion of miR-6867 in ESCC tissues was significantly lower than that in corresponding normal tissues (P<0.05), and its expression was closely correlated with lymph node metastasis and TNM staging (P<0.05). The relative expres-sion of RAPGEFL1 in ESCC tissues was significantly lower than that in corresponding normal tissues (P<0.05), and its expression was closely correlated with lymph node metastasis, pathological differentiation and TNM staging (P<0.05). The expression of miR-6867 and RAPGEFL1 was positively correlated in ESCC tissues (P<0.05). After 5-Aza-dC treatment, the relative expression of miR-6867 and RAPGEFL1 in four cell lines was increased, and the methylation status of RAPGEFL1 was significantly decreased. The methylation rate of RAPGEFL1 in ESCC tissues was significantly higher than that in corresponding normal tissues (P<0.05), which was closely correlated with lymph node metastasis, pathological differentiation and TNM staging (P<0.05). Conclusions: :The occurrence and development of ESCC may be related to the abnormally decreased expression of miR-6867 and RAPGEFL1 and the high methylation status of RAPGEFL1; the expression of miR-6867 was consistent with RAPGEFL1 expression.Moreover, the methylation of RAPGEFL1 gene promoter might be one of the mechanisms underlying the silence of miR-6867 and RAPGEFL1.