Inhibition of U251 human glioma cell line proliferation by knocking down of miR-21 expression in vitro

Abstract

Objective To study the inhibition of U251 human glioma cell line proliferation by knocking down of miR-21 expression in vitro and the possible mechanism.Methods Oligofectamine was used to transfect miRNA-21 antisense oligonucleotides to knock down the miR-21 expression level of U251 human dioma ceill line in vitro.In-situ hybridization and real-time PCR were conducted to detect the miRNA expression of miR-21 anlong different treated groups.The proliferation ability of AS-miR21.scramble and control treated U251 glioma cell were determined by MTr assay.The biological characteristics of U251 cells were evaluated by immunofluoresenee staining and cell flow cytometry.Results The expression level of miR-21 was significantly knocked down bv AS-miR-21 treatment.As MTT assay showed,the tumor cell survival rate was inhihired significantly(F=78.926,P=0.000).G0/G1 phase cell cycle arrest was founded(X2=14.160,P=0.007)and tumor cell apoptosis was induced in As-miR-21 group(F=23341.25,P=0.000).The expression of PCNA.CyelinDi and Bcl-2were down-regulated and mN and Septin-7 were up-regulated iu the AS-miR-21 treated tumor cells.Conclusions The suppressive effect of anti-miR-21 ODNs on the growth of U251 human glioma cell line is significant and miR-21 can be taken as a candidate for gene therapy of human glioma. Key words: miR-21; Antisense oligonuclcotides; U251 human glioma cell line; Gene therapy