Inhibitory efect of Survivin small hairpin RNA on the growth of U251 cells via transferring receptor-mediated deliver

Objective To construct expressing vector of small hairpin RNA (shRNA) targeting Survivin, and investigate the effect of Survivin shRNA on the growth of glioma cell line U251. Methods Oligonucleotides for short hairpin expression targeting Survivin mRNA were inserted into pGCSilencer vector. Recombinant expression vector was identified by enzyme cutting and sequencing analysis. Survivin shRNA was transfected into U251 cells via transferrin-polyethylenimine. Transfected cells were visualized under the inverted fluorescent microscopy and then assayed by the flow cytometry. The expression levels of Survivin mRNA and protein were detected by reverse transcription-polymerase chain reaction (RT-PCR)and Western blotting method respectively. Cytotoxic effects were measured by MTr method. Results The insertion sequence was correctly identified. The RT-PCR and Western blotting method assay revealed that the expression levels of Survivin mRNA and protein in U251 cells were decreased significandy after Survivin shRNA treatment,P ＜0. 05. The growth of U251 cells was significantly inhibited at the 48th, 72nd and 96th h after the treatment with Survivin shRNA as compared with that after treatment with control shRNA and untreated U251 cells, respectively,P ＜ 0.05. Conclusion Survivin shRNA expression vectors can effectively inhibit the growth of U251 cells. Key words: Small hairpin RNA;  Transferrin receptor;  Survivin;  Cell growth;  Glioma