The Study on the Induction Function of SiRNA Mediated Survivin Gene Silence on Nasopharyngeal Carcin

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  To explore the function of Survivin gene expression in patients with nasopharyngeal carcinoma (NPC) and siRNA on controlling CNE-2 nasopharyngeal carcinoma cell proliferation and apoptosis.(Immunology And Histology Chemistry (IHC), in situ hybridization (ISH) and RT-PCR technique are used to detect Survivin protein and mRNA expression; the design software provided by American Ambion Company is used to design RNA(siRNA) sequence which will distract humans Survivin gene; then SilencerTM siRNA Construction Kit is used to synthetize siRNA; Liposome method is used to transfer siRNA into CNE-2 cell strain.RT-PCR technique is used to detect the function of siRNA on controlling Survivin gene expression; MTT assay method is used to test the function of siRNA on controlling the growth and proliferation of cells; FCM method is used to detect the induction function of siRNA on cell apoptosis; electron microscope is used to observe ultrastructure change of CNE-2 cell; Western blotting method is used to detect Survivin gene closed effect.results show that the Survivin protein positive expression rate of nasopharyngeal tissue of NPC is 71.9% while mRNA positive expression rate is 65.6%, the relative expression of mRNA is 4.16× 10-2; data of the control group are 23.3%, 33.3% and 4.42 × 10-4 respectively.For the transfection group with three different siRNA sequences, the CNE-2 cell Survivin mRNA expression level has decreased at different degrees; cell proliferation inhibition rate and apoptosis rate in certain dose have increased along with concentration; siRNA3 transfection group with different doses can induce CNE-2 cell apoptosis at different degrees.Western blotting technique and IHC technique has verified that Survivin protein expression decreases effect and electron microscope has confirmed that there are ultrastructure changes happened in cancer cells.Survivin gene expression of NPC presents tread of increase; Transcription in vitro synthetic siRNA can effectively decrease CNE-2 Survivin gene expression and different sequences of siRNA have different ability of decreasing CNE-2 gene expression; Transfected siRNA3 can effectively restrain the CNE-2 cell proliferation and induce its apoptosis; siRNA is capable of inducing gene silence and providing new perspective for treating NPC.
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