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目的 :检测微RNA-203b(microRNA-203b,miR-203b)基因在食管鳞状细胞癌(esophageal squamous cell carcinoma,ESCC)中的表达及其甲基化状态,探讨miR-203b基因在ESCC发生和发展中的作用。方法 :应用实时荧光定量PCR法检测DNA甲基化转移酶抑制剂5-氮杂-2’-脱氧胞苷(5-aza-2’-deoxycitydine,5-aza-dC)处理前后食管癌细胞系(TE1、TE13、YES-2、EC109和T.TN)和54例ESCC及其癌旁正常组织中miR-203b基因的表达水平。应用甲基化特异性PCR法检测5-aza-dC处理前后5种食管癌细胞系和83例ESCC及其癌旁正常组织中miR-203b基因的甲基化状态。统计学分析ESCC组织中miR-203b基因表达水平和甲基化状态与患者临床病理特征的关系,以及miR-203b基因表达水平与其甲基化状态的相关性。结果 :未经5-aza-dC处理的5种食管癌细胞系中miR-203b基因的表达水平均相对较低,同时表现为高甲基化状态;经5-aza-dC处理后,食管癌细胞系中miR-203b基因的表达水平均升高(P<0.05),同时甲基化程度均降低(P<0.05)。miR-203b基因在ESCC组织中的表达水平明显低于癌旁正常组织(P<0.05),并与肿瘤组织的分化程度密切相关(P<0.05)。ESCC组织中miR-203b基因的启动子区甲基化率明显高于癌旁正常组织(P<0.05),并与肿瘤组织的分化程度密切相关(P<0.05)。发生miR-203b基因甲基化的ESCC组织中miR-203b基因的表达水平明显低于未发生甲基化的ESCC组织(P<0.05)。结论 :miR-203b基因在ESCC中异常低表达,可能与ESCC的发生和发展密切相关,且其启动子区甲基化可能是导致miR-203b基因沉默的机制之一。
Objective: To detect the expression and methylation status of microRNA-203b (miR-203b) gene in esophageal squamous cell carcinoma (ESCC) and to explore the role of miR-203b in ESCC and The role of development. Methods: Real-time fluorescence quantitative PCR was used to detect the expression of 5-aza-2'-deoxycitydine (5-aza-dC) (TE1, TE13, YES-2, EC109 and T.TN) and 54 cases of ESCC and its adjacent normal tissue miR-203b gene expression levels. Methylation-specific PCR was used to detect the methylation status of miR-203b in 5 esophageal cancer cell lines before and after 5-aza-dC treatment and 83 ESCC and its adjacent normal tissues. Statistical analysis of miR-203b gene expression and methylation status in ESCC patients with clinicopathological features, as well as miR-203b gene expression and its methylation status. Results: The expression levels of miR-203b gene in 5 esophageal cancer cell lines without 5-aza-dC were relatively low and hypermethylated at the same time. After 5-aza-dC treatment, esophageal cancer cell lines MiR-203b gene expression levels were increased (P <0.05), while the degree of methylation were reduced (P <0.05). The expression level of miR-203b in ESCC tissues was significantly lower than that in adjacent normal tissues (P <0.05), which was closely related to the differentiation of tumor tissues (P <0.05). The methylation rate of miR-203b gene in ESCC was significantly higher than that in adjacent normal tissues (P <0.05), which was closely related to the differentiation degree of tumor tissues (P <0.05). The expression level of miR-203b in ESCC tissues with methylation of miR-203b gene was significantly lower than that in ESCC tissues without methylation (P <0.05). CONCLUSION: The abnormally low expression of miR-203b in ESCC may be closely related to the occurrence and development of ESCC, and its promoter methylation may be one of the mechanisms leading to the silencing of miR-203b.