摘要
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 （rniR-203b） gene in esophageal squamous cell carcinoma （ESCC）, and to investigate the role of miR-203b in the occurrence and development of ESCC. Methods： The expressions of miR-203b gene in esophageal cancer cell lines including TEl, TEl3, Yes-2, Ec109 and T.TN treated or untreated with DNA methyltransferase inhibitor 5-aza-2＇-detoxycytidine （5-aza-dC） and 54 ESCC tissue specimens and the corresponding non- cancerous tissue specimens were detected by real-time fuorescence quantitative-PCR （RFQ-PCR）. The methylation status of miR-203b gene in five esophageal cancer cell lines treated or untreated with 5-aza-dC and 83 paired ESCC and the corresponding non-cancerous tissue specimens was detected by methylation- specific PCR （MSP）. The relationship between the expression level and methylation status of miR-203b gene in ESCC tissues and the clinicopathologic features of ESCC patients and the correlation of expression and methylation status of miR-203b gene were analyzed by SPSS 13.0 software package. Results： The relatively low expression and the hypermethylation of miR-203b gene were detectable in the five esophageal cancer cell lines untreated with 5-aza-dC. After treatment with 5-aza-dC, the expression of miR-203b was significantly enhanced （P 〈 0.05）, and the methylation level of miR-203b gene was decreased （P 〈 0.05） in esophageal cancer cell lines. The expression of miR-203b in ESCC tissues was significantly reduced as compared with that in the corresponding non-cancerous tissues （P 〈 0.05）, and it was closely associated with the pathological differentiation of ESCC （P 〈 0.05）. The methylation frequency of miR-203b gene promoter in ESCC tissues was significantly higher than that in the corresponding normal tissues （P 〈 0.05）, and it was also associated with the pathological differentiation of ESCC （P 〈 0.05）. The expression of miR- 203b in ESCC tissues with miR-203b gene me
出处
《肿瘤》
CAS
CSCD
北大核心
2014年第9期843-849,共7页
Tumor
基金
国家自然科学基金资助项目(编号:81101854)
