胶质瘤中miR-34a靶向作用于MAPK13 mRNA并抑制其表达

目的预测并验证miR-34a与促分裂原活化蛋白激酶13（MAPK13）的靶向作用关系，寻找胶质瘤基因治疗的新方法。方法将miR-34a相似物转染八胶质瘤细胞A172中，实时定量PCR检测miR－34a在胶质瘤中的表达。目的基因的mRNA水平用实时定量PCR来评估，蛋白水平分别用荧光素酶试验和蛋白印迹来评估。miR－34a与目的基因3′非翻译区（3′UTR）的结合用MAPK13报告实验确认。结果miR－34a相似物转染入胶质瘤细胞A172后在A172细胞中大量表达，实时定量PCR、蛋白印迹和免疫荧光证明miR－34a降低MAPK13的表达量，MAPK13荧光素酶报告实验显示，MAPK13荧光素酶活性被miR－34a降低。结论miR－34a通过与MAPK133′UTR区的特异结合作用发挥对胶质瘤责任基因MAPK13的靶向沉默作用，可能成为未来胶质瘤基因治疗的新选择。

MiR-34a directly targets mRNA of MAPK13 and inhibits its expression in glioma

Objctive To predict and verify whether miR-34a directly targerts mitogen-activated protein kinase 13 （MAPK13） and to find new ways for glioma gene therapy. Methods Human glioma cell line A172 was transfected with miR-34a mimics. Real-time PCR was used to detect its expression in glioma cell. Potential target gene expression was assessed by Western bl0t and immunofluorescence for proteins, and by quantitative real-time PCR for mRNAs. The binding between miR-34a and 3′ UTR of MAPK13 was validated using 3′ UTR reporter assay. Results After transfecting miR-34a mimics into human glioma cell line A172, miR-34 was largely expressed in A172. Real-time PCR, immunofluorescence and Wstern blot showed that miR-34a reduced the expression of target gene MAPK13. 3′ UTR reporter assay showed that the luciferase activity of MAPK13 was inhibited by naiR-34a. Conclusion MiR-34a silencing MAPK13 by directly binding to 3′ UTR of MAPK13 may become a new strategy for future glioma gene therapy.