Inhibition of the expression of VEGF gene in nasopharyngeal carcinoma cells by microRNA.
- Author:
Xinzhang CAI
1
;
Wei WEI
;
Suping ZHAO
;
Yaoyun TANG
;
Chufeng HE
;
Chenglong WANG
Author Information
1. Department of Otolaryngology-Head and Neck Surgery, Xiangya Hospital, Central South University, Changsha, 410008, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Cell Line, Tumor;
Female;
Genetic Therapy;
Humans;
Mice;
Mice, Inbred BALB C;
Mice, Nude;
MicroRNAs;
genetics;
Nasopharyngeal Neoplasms;
genetics;
metabolism;
Plasmids;
Vascular Endothelial Growth Factors;
genetics;
metabolism
- From:
Journal of Clinical Otorhinolaryngology Head and Neck Surgery
2010;24(15):703-707
- CountryChina
- Language:Chinese
-
Abstract:
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.
