Effect of Survivin targeting on cell proliferation and apoptosis in glioma cell line

Objective To investigate the effects of antisense Survivin-Lipcompound on U251 human glioma cell proliferation and apoptosis. Methods Survivin oligonucleotide (ODN) was transfected into the U251 glioma cells mediated by Lip reagent. The expression of Survivin protein was detected by Western blotting. MTT assay was applied to determine the proliferation of U251 cells. Apoptosis rate and mitochondrial membrance potential (MMP) were evaluated by flow cytometric analysis. The morphological changes were assessed by an electron microscope. Laser scanning confocal microscope immunofluo-rescence analysis was performed to detect the sub-cellular localization of Survivin protein on treated cells and untreated cells. Results Antisense compound efficiently down regulated the Surviving expression. As revealed by gradually increased apoptosis rate and decreased MMP in a time dependent manner, and their morphological changes, treatment with antisense compound induced apoptosis and inhibited cell growth. Fluororescein isothiocyanate (FITC)-labeled immunofluorescence staining of Survivin clearly showed that Survivin was expressed mainly in the formation of a spotted distribution inside the cytoplasm of untreated cells. Survivin protein molecules were clearly seen in the cytoplasm of the untransfected cells and distributed like spots and almost disappeared in the transfected cells with morphological changes conforming to the changes of apoptosis. Conclusion Survivin protein is a key molecule connecting proliferation with apoptosis and antisense oligonucleotides targeting Survivin has a bright prospect in the treatment of glioma cells.