胶质瘤中miR-410靶向作用于MET的初步研究

Preliminary study of mi R-410 targeting on MET in gliomas

目的探讨mi R-410在胶质瘤细胞的作用机制。方法收集50例人胶质瘤标本以及胶质母细胞瘤U251和U87细胞系。采用mi Randa靶基因预测软件对mi R-410的靶基因预测分析,初步判断其与MET基因的相关性;利用免疫组化和原位杂交技术检测人脑胶质瘤标本中mi R-410和MET的表达;荧光素酶报告基因实验检测mi R-410对MET的靶向作用;Western blot检测mi R-410模拟物转染肿瘤细胞后MET蛋白的变化。结果经mi Randa靶基因分析:MET m RNA的3’非编码区(3’UTR)与mi R-410的种子序列存在理论上的互补匹配序列。免疫组化和原位杂交技术检测:肿瘤标本中mi R-410和MET具有负相关性(r=-0.69783,P<0.001)。荧光素酶实验进一步证实胶质母细胞瘤中MET基因是mi R-410的直接靶点。肿瘤细胞转染mi R-410模拟物后对MET蛋白的表达具有明显的抑制作用(P<0.05)。结论在胶质瘤细胞中mi R-410靶向作用MET,可能是胶质瘤治疗的新靶点。

Objective To explore the functional mechanism of miR-410 in glioma cells.Methods Fifty human glioma samples and glioblastoma cell lines U251 and U87 were collected.The genetic analysis of correlation of miR-410 and MET gene was judged by the miRanda target gene prediction software.Immunohistochemistry and in situ hybridization technique were used to confirm the expressions of miR-410 and MET in the human brain glioma samples.Further,Luciferase experimental method was used to test the targeting effect of miR-410 on MET.Western blot was used to detect the change of MET protein after the transfection of miR-410 analogue into U87 and U251 tumor cells.Results The matching sequences between 3＇untranslated region（3＇UTR） of MET m RNA and seed sequences of miR-410 existed theoretically,which were confirmed by miRanda target gene analysis.The negative correlation of miR-410 and MET in tumor samples was confirmed by immunohistochemistry and in situ hybridization（r =-0.69783,P〈0.001）.Luciferase experiments further confirmed that MET gene was the direct target of miR-410 in human glioblastoma.After the tumor cells were transfected with miR-410 simulation,the MET expression was inhibited significantly（P〈0.05）.Conclusion miR-410 targets on MET in gliomas,and may be the new target for glioma treatment.