The Effect Of MiR-21 On Glioma Cells (U373MG) Growth And Identification Of Its Target

Objective:MicroRNAs(miR-21) are small noncoding RNA sequence.As a molecule, they generally exist in adjusting gene expression.MiRNAs involve in many important proceeding,There are many researches indicate that miRNAs may have potential relation with cancer.In different cancer cell lines of different tissues,the level and the regulation effect of miRNAs are different.The study predict and confirm the target genes of miR-21 in human tumor cells by miRNA microarray and reporter assay technologies,and deeply study the biological functions which miR-214 involved through regulating the expression of target genes..It may provid some information to comprehend the effect of miR-21 in tumor formation.Methods:Four antisense oligonucleotides of miRNAs were transfected to human brain glioblastoma U373MG cells by Lipofectamine,and gapas control as control.After 48 h,the activity of transfected cells was determined by MTT.The changes of genes expression table of human brain glioblastoma U373MG cells transfected miR-21 ASO and gapas control are checked by cDNA assay.The target gene of miR-21 are found thought bioinformatic technique and the results of cDNA assay,constructed pCDNA₃/ EGFP-LRRFIP1 3'UTR vector.It is validated that LRRFIP1 is the target gene of miR-21by the express of GFP report vector.Results: 1.After transfected miR-21 ASO,the cell activity of human brain glioblastoma U373MG cell is markly decrease(P＜0.05).There are not differences in cell activity among other groups.2.The result of cDNA microarray shows that 179 genes up-regulate and 153 genes down-regulate.These genes can be divided into 5 functional categories,which including apoptosis,cancer,signal transduction,liver enzyme metabolism,and cytokine related genes.3.We choose some genes.These genes were confirmed by semi-quantitative RT-PCR.Up-regulation genes have LRRFIP1,SHB,PTK2,FLII,MYLK,HDAC1,AKT1,PTPRF,CDB1,ITGB4,CLU,PPP2R1A,Down regulation genes have PDCD5,STK6,GADD45B,YY1.4.LRRFIP1 may be the target gene of miR-21 thought bioinformatic technique and validated by the express of GFP report vector.Conclusions:As a antiapoptosis factor in human brain glioblastoma U373MG cell the function of microRNA21 is inhibited through transfer its anti-sense oligonucleotides, the human brain glioblastoma U373MG cell activity is markly decrease.Some genes of apotosis and metabolism up-regulate or down-regulate,this may have relate with adjusting cancer cell achieve a new balance.The study predict and confirm that the target genes of miR-21 in human tumor cells may be LRRFIP1 by miRNA microarray and reporter assay technologies...