The Study Of The Relationship Between The Expression Of Mir-126 And Cell Proliferation, Invasion In Glioma

Objective: To explore mir-126 expression in human gliomas and its correlation with malignant glioma U87 cell proliferation,invasion, and to further define the mechanism of mir-126 in gliomas.Methods:â‘ Real-time fluorescence quantitative PCR was used to determine the expression of mir-126 in 24 cases of glioma tissues pathologically confirmed and 6 cases of normal brain tissues respectively.â‘¡Computer-designed oligo nucleotide sequences which express the pre-miRNA were connected to plasmid pcDNA6.2- GW/EmGFP-miR after annealing, then the recombinant plasmids would fulfill the sequences analysis. Fluoromicroscopy was applied to observe the transfectional effect after the plasmids enter U87 cells via lipofectamine. The quantity of mir-126 gene expression could be detected by RT-PCR and Gel electrophoresis.â‘¢MTT method was used to analysis the proliferation of U87 glioma cells after mir-126 transfection.â‘£The relative expression of MMP-2 and MMP-9 gene was quantified by RT-PCR, the expression of MMP-2 and MMP-9 protein was detected by western blot, and cell invision was assessed by Transwell chamber assay.Results:â‘ Real-time quantitative PCR results showed that the expression of mir-126 in glioma tissues lower than in normal brain tissues.â‘¡Green fluorescence could be observed in post-transfectional U87 cells,which convinced that cell transfcetions were successful,and transfected plasmids were expressed in cells. RT-PCR and Gel electrophoresis results showed that mir-126 expression would increase after transfection.â‘¢Mir-126 inhibited the proliferation of U87 cells.â‘£Mir-126 inhibited the activity of MMP-2,MMP-9 and the invision of human U87 glioma cells. Conclusion: This study suggest that the expression of mir-126 in glioma tissues lower than in normal brain tissue. MicroRNA recombinant plasmid is constructed successfully, and comfirmed its inhibition of the proliferation and invision of malignant glioma U87 cells.