Abstract
Background: To explore the impact of pU6-based tandem survivin and CDK1-specific short hairpin RNA on the biological behaviors of CNE-2 nasopharyngeal carcinoma cells in vitro and in vivo. Patients and Methods: The vectors of pU6-survivinshRNA, pU6-CDK1shRNA and pU6-survivinshRNA-CDK1shRNA were constructed and transfected into CNE-2 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 CNE-2 cells, and flow cytometry was performed to determine the apoptosis of CNE-2 cells. The effects of interfering survivin and CDK1 on tumorigenesis were evaluated by tumor xenografts experiments. Results: Effective plasmids were successfully constructed knocking down survivin and/or CDK1. The proliferation inhibition of CNE-2 cells by pU6-survivinshRNA-CDK1shRNA (32.5%) was higher than that of by pU6-survivinshRNA (25.6%) and pU6-CDK1shRNA (15.6%), and apoptosis in CNE-2 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 more effective than interfering alone component in inhibiting tumor growth of fBalb/C nude mice xenografted with CNE-2 cells. Conclusion: The results altogether indicate that interfering survivin and CDK1simutaneously can produce synergistic effects of anti-nasopharyngeal carcinoma, which could be a potential therapeutic method.
Highlights
Nasopharyngeal carcinoma (NPC) is one of the most popular cancers in southern China [1]
Simultaneously interfering survivin and CDK1 is more effective than interfering alone component in inhibiting tumor growth of fBalb/C nude mice xenografted with CNE-2 cells
The five year survival rate is only 50% - 60% due to factors below: most NPC organization cells is undifferentiated with high malignancy; the NPC anatomical location impedes radical resection and the remaining NPC cells serve as seeds for future recurrence and metastasis in future; NPC cells are insensitive to chemotherapy; NPC cells develop radio resistance during radiotherapy
Summary
Nasopharyngeal carcinoma (NPC) is one of the most popular cancers in southern China [1]. It is essential to develop a new therapeutic technology to improve the unsatisfactory outcome of NPC treatment. With the development of technology of gene recombination and transgenosis, RNA interference technology has become a powerful tool of knocking out targeted gene expression, which offers an optimistic treatment for various cancers. Survivin is highly expressed in almost all cancers, including NPC. Expression of survivin is undetectable or rather low in almost all normal mature cells, which makes survivin an ideal target for various cancers [3] [4] [5] [6]. Overexpression of CDK1 in cancers induces cell proliferation and chromosomal instability, which makes CDK1 a potential therapeutic target [10] [11] [12]. The study is aimed at observing impacts of knocking out survivin and CDK1 simultaneously on nasopharyngeal carcinoma by constructing pU6-survivinshRNA-CDK1shRNA, and evaluating the potential value of survivin and CDK1 in tumor therapy
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