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To explore the impact of pU6-based tandem survivin and CDK1-specific short hairpin RNA on the biological behaviors of CNE2 nasopharyngeal carcinoma cells in vitro and in vivo.The vectors of pU6-survivinshRNA, pU6-CDK1shRNA and pU6-survivinshRNA-CDK1shRNA were constructed and transfected into CNE2 cells with Lipofectamine TM 2000, respectively.The mRNAs and proteins of CDK1 and survivin were determined by RT-PCR and Western blotting, accordingly.MTT assay was employed to evaluate the proliferation of CNE2 cells, and flow cytometry performed to determine the apoptosis of CNE2 cells.The effects of interfering survivin and CDK1 on tumorigenesis were evaluated by tumor xenografls experiments.Effective plasmids knocking down suvrvin and/or CDK1 were successfully constructed.The proliferation inhibition of CNE2 cells interfered by pU6-survivinshRNA-CDK1shRNA (32.5%) was higher than those interfered by pU6-survivinsshRNA(25.6%) or pU6-CDK1shRNA (15.6%), and apoptosis in CNE2 cells simultaneously interfering survivin and CDK1 (15.2%) dramatically increased when compared to those of interfering survivin (5.4%) or CDK1 (4.7%) alone.Furthermore, simultaneously interfering survivin and CDK1 is superior to interfering either component in inhibiting tumor growth in Balb/C nude mice xenografted with CNE2 cells.These findings altogether indicate that simultaneously interfering survivin and CDK1 can produce synergistic effects of anti-nasopharyngeal carcinoma, which may be an attractive therapeutic method.