Knock-down of IKKε inhibits proliferation and invasion of U251 glioma cells in vitro

Abstract

Objective To investigate the effect of IKKe knock-down on the biological characteristics of U251 glioblastoma cells. Methods IKKe small interfering RNA (IKKe siRNA) mediated by lipofectamine were transfected into U251 cells while cells transfected with scrambled siRNA and control cells were prepared.RT-PCR was employed to detect expressions of IKKe in the transfected cells.The cell proliferation was determined by MTT assay.Flow cytometry was used to monitor changes in cell cycle.Cell invasion was evaluated by Transwell assay.Moreover,the proliferating cell nuclear antigen (PCNA),cyclin D 1 and matrix metalloproteinase-9 (MMP9) that regulated proliferation,invasion and cycle progression of the transfected cells and the changes of NF-κB after transfection were examined by Western blotting. Results RT-PCR revealed that the proliferation of U251 cells was inhibited,the cell cycle was arrested in G0/G1 phase and the invasive activity was attenuated in cells transfected with IKKe siRNA,with significant differences as compared with the cells transfected with scrambled siRNA and control cells (P＜0.05).The expressions ofPCNA,MMP9 and cyclin D1 were down-regulated in the IKKe knock-down cells, as compared with the other 2 groups. In addtion, transposition of NF-κB reduced from the cytoplasm to the nucleus after transfection. Conclusion As IKKe plays a vital role in proliferation and invasion of glioma cells,it may serve as a potential target ofgene therapy for glioma. Key words: IKKe; Glioma; Proliferation; Invasion; Cell cycle