Effects Of RNAi-Silenced Livin Gene On Glioma Cell Lines U251

Objective:To construct eukaryotic expression vectors by using the pSilencer2. 0 vector for inhibiting human livin gene by RNA interference, and to detect the effect of the silenced livin gene on the growth and apoptosis of U251 cells.Methods:Two target gene segments were synthesized and cloned into the pSilencer2. 0 vector respectively to construct two recombinant eukaryotic expression vectors: pSilencer2.0-liv1 and pSilencer2.0-liv2, both of which were identified by enzyme digestion analysis. Then the U251 cells were transfected with the recombinant vectors and the interfering effects were detected by RT-PCR and Western blot. The proliferation of the U251 cells was detected by MTT method and their apoptosis index detected by flow cytometry.Results:The results of Enzyme digestion analysis showed that two target segments were cloned into pSilencer2. 0 vectors. The results of RT-PCR and Western blot indicated that pSilencer2.0-liv1 and pSilencer2.0-liv2 vectors could knock down the transcription and expression of livin gene, but the effect of pSilencer2.0-liv2 vectors was better than that of the pSilencer2.0-liv1 vectors. Obvious decrease in growth of the U251 cells was observed after transfected with pSilencer2.0-liv1 and pSilencer2.0-liv2 vectors, along with 17.7% increase of the apoptosis index in the U251 cells transfected with pSilencer2.0-liv2 vectors.Conclusion : The transcription and expression of livin gene were inhibited effectively in the glioma cell line U251. After transfected with pSilencer2.0-liv2 vectors, the apoptosis index of the U251 cells was increased and the growth inhibited.