Mi-34a Affects The Invasion And Metastasis Of Glioma Cells Through Its Target Gene MMP2

Objective: To investigate the effect of miR-34 a on the invasion and migration of glioma cells through the target gene MMP2 by detecting the expression of miR-34 a and MMP2 in glioma cell lines.Methods: The expression of MMP2 in glioma cell lines was detected by real-time PCR and miR-34 a.U87 and A172 were selected and miR-34 a mimics and miR-34 a inhibitors were used.Group transfection,miR-34 a was observed by qRT-PCR,qRT-PCR and western blot were used to observe the expression level of MMP2;mRNA database was used to predict miR-34 a binding to MMP2 region,and then wild type and mutant MMP2 3’UTR fluorescein were constructed.The enzyme plasmid and the miR-34 a plasmid were tested by fluorescent Samui reporter gene;the invasion and migration ability of U87 cells were observed by Transwell invasion and scratch test.Results:(1)The content of MMP2 protein in each glioma cell line was highly expressed,among which U87 and A172 were most obvious,and the mRNA expression level of MMP2 was basically the same as that of protein,but the expression level of miR-34 a mRNA was relatively low.The mRNA expression level of miR-34 a was negatively correlated with MMP2,and MMP2 might be a downstream target gene of miR-34 a.(2)After miR-34 a mimic and inhibitor were transfected into U87 and A172 cells,the protein and mRNA levels of MMP2 transfected into mimic cell line were significantly decreased,and the protein and mRNA levels of MMP2 in transfected inhibitor cell lines were significantly decreased.Raise.(3)The luciferase plasmid of the wild type and mutant MMP2 3’ UTR region and pcDNA5-miR-34 a plasmid were transferred into U87 and A172 cells respectively,and the luciferase activity and blank of the mutant MMP2 3’UTR region in the experimental group.There was no significant difference in the control group,but the luciferase activity of the wild-type MMP2 3’UTR region in the experimental group was significantly lower than that in the blank control group.(4)Cell invasion and migration assays by Transwell invasion assay and scratch assay showed that miR-34 a inhibited tumor invasion and migration,and reversed the ability of MMP2 to enhance invasion and migration.Conclusion:(1)The mRNA expression level of miR-34 a is negatively correlated with the mRANA expression of MMP2,and MMP2 may be the downstream target gene of miR-34 a.(2)miR-34 a in the glioma cell line can down-regulate the protein and mRNA expression levels of MMP2 by binding to the specific sequence of the 3’ UTR region of MMP2.(3)miR-34 a is a tumor suppressor gene in glioma,which inhibits the invasion and migration of glioma cells by down-regulating the expression of MMP2.