人脑胶质瘤中miR-106b~25基因簇表达的研究

  目的  检测胶质瘤细胞系及组织中miR-106b~25基因簇成员miR-106b、miR-93、miR-25的表达情况。  方法  运用实时定量PCR的方法检测不同人脑胶质母细胞瘤细胞系及不同病理级别胶质瘤标本中miR-106b~25基因簇成员miR-106b、miR-93和miR-25的表达水平。原位杂交技术检测含不同病理级别及瘤旁正常脑组织的胶质瘤组织芯片中miR-106b~25基因簇成员的表达情况，进而分析该簇miRNAs的表达与胶质瘤病理级别的相关性。  结果  以正常脑组织表达量为基准，3种miRNA在所有检测细胞系中均有增高的趋势。收集43例胶质瘤标本中，RT-PCR结果显示，随着肿瘤病理级别的升高，3种miRNA在各组中的平均表达量也逐步升高。其中，miR-106b（F=4.479，P=0.018）和miR-93（F=3.493，P=0.040）各组间的表达差异均有统计学意义。而miR-25表达无明显的统计学差异（F=2.766，P=0.075）。原位杂交结果显示3种miRNA的表达在高级别胶质瘤中的表达量明显高于低级别肿瘤。Spearman等级相关分析表明3种miRNA的表达信号强度分布均与胶质瘤的WHO病理分级呈正相关，在miR-106b、-93、-25中的相关系数分别为r_s=0.617（P＜0.001）、r_s=0.438（P＜0.001）、r_s=0.463（P＜40.001）。  结论  miR-106b~25在胶质瘤中表达呈不同程度增高，并与肿瘤分级呈正相关。 

A study on the expression of miR-106b~25 cluster in human glioma

  Objective  To detect the expression of miR-106b~25 cluster in glioma cell line and tissues.  Methods  Real-time PCR was performed to determine the expression of miR-106b~25 cluster members (miR-106b, miR-93, and miR-25) in different human glioblastoma cell lines. Different pathological grade glioma specimens were surgically removed. In-situ hybridization was performed to detect the expression of miR-106b~25 cluster members in different pathological levels of glioma tissues.  Results  In the expression of the benchmark on normal brain tissues, three kinds of miRNAs in all test cell lines have a tendency to increase. Based on the expression of the pathological level Ⅰ average rate in 43 cases of glioma specimens collected after neurosurgical operations, the real-time PCR results showed that the average expression quantity of the three kinds of miRNAs in each group gradually increase. The increase in tumor pathological levels results in statistically significant expression differences of miR-106b and miR-93 between the groups (F=4.479, P= 0.018 and F=3.493, P=0.040, respectively). However, miR-25 expression differences between the groups have no statistically significant differences (F=2.766, P=0.075). In situ hybridization results show that the expressions of three miRNAs in high grade gliomas are significantly higher than that in the low-level tumor. Spearman rank correlation analysis results indicate that the expression of these miRNAs signal-intensity distribution is positively correlated with glioma, in accordance with WHO pathology classification. The correlation coefficient for miR-106b, miR-93, and miR-25 are 0.617, 0.438, and 0.463, respectively (P < 0.001).  Conclusion  The expression of miR-106b~25 cluster members is up-regulated in the glioma and is positively correlated with tumor grade.