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目的:检测miR-129成熟体miR-129-5p在人脑胶质瘤组织中的表达特点,探讨人脑胶质瘤组织中miR-129-2表达异常是否与miR-129-2启动子甲基化有关。方法:收集湘雅医院神经外科2013年10-12月收治的胶质瘤患者肿瘤组织21例、脑外伤患者非肿瘤脑组织标本21例。Real-time PCR检测脑胶质瘤组织和非肿瘤脑组织、脑胶质瘤细胞株和正常脑胶质细胞株中miR-129-5p的表达;甲基化特异性PCR(methylation-specific PCR,MSP)检测胶质瘤组织中miR-129-2基因启动子甲基化的情况;亚硫酸氢盐硫化测序PCR(bisulfite-sequencing PCR,BSP)分析经甲基化转移酶抑制剂5-氮杂-2’-脱氧胞苷(5-aza-2’-deoxycitydine,5-Aza-d C)处理后的胶质瘤细胞miR-129-2基因启动子甲基化水平的变化,同时Real-time PCR检测miR-129-5p的表达变化。结果:miR-129-5p在人脑胶质瘤组织和细胞中表达显著低于非肿瘤脑组织和正常脑胶质细胞(P<0.05),胶质瘤组织中miR-129-2基因的启动子甲基化率显著高于正常脑组织(P<0.05),使用5-Aza-d C去甲基化处理脑胶质瘤细胞后miR-129-2基因的启动子甲基化程度降低、miR-129-5p表达显著升高(P<0.05)。结论:miR-129-5p在脑胶质瘤中显著低表达,miR-129-2启动子甲基化是导致其低表达的机制之一。
AIM: To detect the expression of miR-129-5p in human glioma tissue and to investigate whether the abnormal expression of miR-129-2 in human glioma is associated with miR-129-2 promoter Based on. Methods: Twenty-one cases of glioma were collected from neurosurgical department of Xiangya Hospital from January to December in 2013, and 21 cases of non-tumor brain tissue from patients with traumatic brain injury. Real-time PCR was used to detect the expression of miR-129-5p in glioma tissues, non-tumor brain tissues, glioma cell lines and normal glia cell lines. Methylation-specific PCR MSP) was used to detect the promoter methylation of miR-129-2 gene in glioma tissue. Bisulfite-sequencing PCR (BSP) was used to analyze the methylation of the promoter of methylated transferase 5-aza -2-deoxycytidine (5-Aza-dC) treatment of glioma cells miR-129-2 gene promoter methylation levels, while Real-time PCR detection of miR-129-5p expression changes. Results: The expression of miR-129-5p in human glioma tissues and cells was significantly lower than that in non-tumor brain tissues and normal glial cells (P <0.05), and the activation of miR-129-2 in glioma tissues The methylation rate of miR-129-2 gene in 5-Aza-d C-demethylated glioma cells was significantly lower than that in normal brain tissue (P <0.05) miR-129-5p expression was significantly increased (P <0.05). Conclusion: miR-129-5p is significantly down-regulated in gliomas, and the methylation of miR-129-2 promoter is one of the mechanisms leading to its low expression.