Objective To analyze miR-10b expression level and the gene upstream methylation level in schwannomas so as to explore and identify the potential target genes for miR-10b in schwannomas .Methods The miR-10b with its potential target genes including HOXB 3 ,HOXD10 ,PTEN ,PIK3CA ,MAPRE1 and HADC4 were quantitatively analyzed by PCR in 13 cases of schwannomas and 6 cases of human vestibulocochlear nerves . We studied the correlation between the differentially expressed genes and the clinical characteristics of schwannomas . Finally ,the differences in miR-10b gene upstream methylation levels were measured and analyzed by pyrosequencing between schwannomas and normal vestibulocochlear nerves .Results Compared with that of normal nerves ,the expression level of miR-10b was significantly higher (P=0 .0003) while the level of PTEN was lower (P=0 .0047) in schwannomas .Negative correlation existed between the levels of miR-10b and PTEN (P=0 .001 , r= -0 .689) . Moreover ,the methylation level of the miR-10b gene promoter was downregulated in schwannomas ;it had negative correlation with the expression level of miR-10b (P= 0 .011 , r= -0 .571) .There was a significant difference in tumor mass diameter between miR-10b higher expression group and lower group (P=0 .016);however ,there was no difference in age or recurrence rate (P>0 .05) .Conclusion The downregulation of methylation level of the promoter leads to higher expression of miR-10b gene ,and it may targetedly inhibit the expression of PTEN .%目的 分析miR-10b小RNA在神经鞘瘤中的表达水平和该基因上游甲基化水平,并识别miR-10b在神经鞘瘤中的潜在靶基因.方法 应用实时定量PCR定量miR-10b及潜在靶基因HOXB3、HOXD10、PT EN、PIK3CA、MAPRE1、HADC4在13例神经鞘瘤和6例正常前庭蜗神经的表达;进一步分析差异表达基因与神经鞘瘤临床特征的相关性;最后应用焦磷酸测序分析两组之间miR-10b基因上游甲基化水平差异.结果 与正常神经相比,神经鞘瘤组织中miR-10b分子表达水平显著升高(P=0.0003),而PT EN表达显著下降(P=0.0047),两者显著负相关(P=0.001,r=-0.689).此外,miR-10b基因启动子区内甲基化水平下调,并与该基因表达显著负相关(P=0.011,r=-0.571).miR-10b高表达组与低表达组比较,肿瘤直径有差异(P=0.016);但发病年龄及1年后肿瘤复发率无差异(P>0.05).结论 神经鞘瘤中miR-10b启动子甲基化水平下降引起该基因持续高表达,而miR-10b可能靶向抑制PT EN表达.
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