Objective To investigate the inhibition effect of RNA interference targeting survivin gene of human nasopharyngeal carcinoma CNE2 cells in vitro and in vivo. Methods pGenesil-1 -survivin expression vector of short hairpin RNA (shRNA) specific to survivin gene was transfected into human nasopharyngeal carcinoma CNE2 cells through Lipofectamine,2000 reagent. Stably transfected CNE2 cells were obtained after G418 screening and detected by fluorescence microscope and flow cytomytry (FCM) to analyze the cell transfection efficiency. The mRNA and protein expression of survivin were determined by reverse transcription-polymerase chain reaction (RT-PCR) and FCM, respectively. Cell proliferation ability was determined by methyl thiazol tetrazolium ( MTT) assay. BALB/c nude mice bearing stably transfected human nasopharyngeal carcinoma CNE2 cells were established. Tumor sizes were measured and tumor growth curves were made. Results FCM results showed that positive transfected percentage of pGenesil-1 -survivin was (87.13 ±.21)%. The mRNA and protein expression of survivin inhibited by 64.55% and 51.24% respectively and deduced significantly when compared with that of either pGenesil-1-NC ( expression vector with scrambled small interfering RNA) transfected cells or control cells (P < 0. 05). The MTT results showed that the proliferation rate of CNE2 cells stably transfeoted with pGenesil-1-survivin was significantly reduced when compared with that of either pGenesil-1-NC group or control group {P <0. 05). In vivo mice bearing tumor, pGenesil-1-survivin showed a significant inhibitory effect on the growth of transplanted human nasopharyngeal carcinoma, resulting in a statistically significant difference compared with the other groups (P <0.05). Conclusion Survivin gene silence can effectively inhibit the proliferation ability of human nasopharyngeal carcinoma CNE2 cells in vitro and in vivo. Survivin gene may be a promising target in the gene therapy of human nasopharyngeal carcinoma.
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维普
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