Objective: To explore the regulative effect of expression of VEGF gene in nasopharyngeal carcinoma, and to discuss the future application of microRNA in the gene therapy for nasopharyngeal carcinoma.
Method: We constructed the recombination miRNA plasmid vectors which target VEGF gene and plasmids were transfected into CNE-2 cells by using Lipofectamine 2000 Reagent. The VEGF mRNA and VEGF protein were detected by reverse transcriptase polymerase chain reaction (RT-PCR) and Western blotting respectively. WST-8 assay was used to determine the inhibitory effect of microRNA on cell growth. Stable cell lines and wild type CNE-2 cell line were inoculated to subcutis of nude mice to establish animal models. The tumor growth and volume were observed.
Result: After the transfection of CNE-2 cells , the expressions of VEGF mRNA and VEGF protein were down-regulated at different degree. Whereas, CNE-2 cell growth showed no change by observation of fluorescence microscopy, and cell proliferation was not inhibited in WST-8 assay. However, in vivo, growth of xenograft was inhibited in preliminary experiments of nude mice.
Conclusion: By miRNA plasmid constructed artificially, miRNA can effectively interfere nasopharyngeal carcinoma cells by down-regulating the expressions of VEGF gene, therefore can inhibit the growth of tumor xenografted in vivo. Future application of microRNA in the gene therapy of nasopharyngeal carcinoma might be expected.