Estabishment Of Stathmin Silencing Nasopharyngeal Carcinoma Cell Line And Explortion Of Its Biological Characteristics

Objective:To construct the stathmin-siRNA recombinant plasmid express-ion vector,build stathmin silence the nasopharyngeal carcinoma5-8F cell lines, study the effects of stathmin silencing on nasopharyngeal carcinoma5-8F cell proliferation,cycle and tumorigenicy,and provide the experiment basis for nasopharyngeal carcinoma gene therapy.Methods:(1) Synthesize stathmin specific siRNA fragment,and the Frag-ment was inserted into pGenesil-1.1plasmid expression vector to construct the recombinant plasmid expression vector.(2)Recombinant plasmid expression ve-ctor was transfected in5-8F cell line, the transfected cells were identify by RT-PCR and western blotting to affirm the stable stathmin expression inhibited5-8F cell lines.(3) Cellular proliferation was tested by MTT experimental analy-sis,cell cycle was tasted by flow cytometric analysis.(4) Cancer cell tumorigeni-city was tested by nude mice experiment.Results:Obtaining stathmin siRNA pGenesil-1.1recombinant plasmid expression vector and stathmin silencing nasopharyngeal carcinoma5-8F cell line. The stathmin-silenced5-8F cell line was cultured for MTT experimental, the results showed the cell proliferation inhibition rate of stathmin-silenced5-8F cells at48h,72h and96h were significantly higher than negative control group (33.67±2.52)%vs(8.13±0.60)%,(28.33±1.15)%vs(3.20±0.53)%,and(31.67±1.53)%vs(3.97±0.15)%,P<0.01.Stathmin-silenced5-8F cells in G2phase was (15.8±0.55)%,showing significance with negative control group (11.3±0.79)%and blank control group(10.9±0.92)%,P<0.05. The transplanted tumor weight of stathmin-silenced5-8F cells was (0.32±0.21)g,significantly lower than that of negative control group (1.10±0.40)g and blank control group(1.52±0.30)g, P<0.05.Conclusion:Stathmin-siRNA plasmid expression vector was successfully constructed, and nasopharyngeal carcinoma cell line of stathmin inhibited expre-ssion is established;Stathmin-silence inhibits malignant phenotype of nasophary-ngeal carcinoma5-8F cell line.