中文摘要：

前期的研究证明,人腺苷酸活化蛋白激酶5（ARK5）通过Gab2-Akt-ARK5通路,促进胶质瘤的侵袭。然而,ARK5的调节机制尚不清楚。本研究旨在探讨miR-424是否通过与ARK5 mRNA结合,进而影响胶质瘤侵袭。通过Targetscan找到与ARK5的3＇-UTR区互补结合的microRNA（miR-424）。运用实时荧光定量PCR（qRT-PCR）检测不同胶质瘤细胞系中miR-424表达水平,发现3种胶质瘤母细胞（高级别胶质瘤细胞）株中,miR-424表达量均不同程度低于低级别胶质瘤H4细胞株。miR-424及miR-424抑制剂质粒转染胶质瘤细胞系H4、LN-229并结合蛋白质印迹法显示,miR-424可负向调控ARK5蛋白表达。qRT-PCR显示,在胶质瘤细胞内过表达miR-424后,ARK5mRNA无显著变化,提示miR-424在翻译水平影响ARK5蛋白表达。Transwell侵袭实验显示,过表达miR-424导致胶质瘤细胞体外侵袭能力明显减弱。双荧光素酶基因报告检测显示,miR-424能与ARK5 mRNA的3＇-UTR结合,抑制荧光素酶活性。上述结果提示,miR-424可以结合ARK5mRNA的3＇-UTR而抑制ARK5蛋白翻译,从而抑制胶质瘤细胞侵袭。

英文摘要：

Previous studies have demonstrated that human adenylate activated protein kinase 5 （ARK5） promotes glioma invasion through the Gab2-Akt-ARK5 pathway, but the regulatory mechanism of ARK5 is unclear. This study was designed to investigate how miR-424 regulates the expression of ARK5 mRNA and affects glioma invasion. MicroRNA-424 （miR-424） was found by Targetscan in the region complementary with the 3＇-UTR region of ARKS. The real-time fluorescence quantitative PCR （quantitative real-time PCR, qRT-PCR） was used to detect the expression levels of miR-424 in different glioma cell lines. We found that miR-424 expression was significantly lower in high grade glioma cell lines than H4 cell line with low invasive ability. H4 and LN-229 were transfected by miR-424 plasmid and miR-424 inhibitor and their respective controls. Western blotting showed that miR-424 negatively regulated the expression of ARK5 protein. QRT-PCR showed that there was not significant change ofARK5 mRNA in glioma cells after overexpression of miR-424, miR-424 affects the expression of ARK5 protein at the translation level. Transwell invasion assay showed that the invasion ability of glioma cells was significantly decreased after overexpression of miR-424. Double luciferase reporter assay showed that miR-424 could inhibit the luciferase activity of mRNA by binding to ARK5 3＇-UTR. These data suggest that miR-424 can inhibit the translation of ARK5 protein by targeting the 3＇-UTR of ARK5 mRNA, thereby inhibiting the invasion of glioma cells.