The construction of eukaryotic expression vector of short hairpin RNA for CXCR4 / 实用口腔医学杂志

ABSTRACT

Objective:To construct of eukaryotic expression vector of RNA interference specific for CXCR4. Methods: Genome sequences of CXCR4 gene were retrieved from Genbank and cDNA was designed coding expression of shRNA (small hairpin RNAs) for CXCR4 gene. The cDNA was synthesized and inserted into plasmid pWH1, and the recombinant pWH1-CXCR4 expression vector was identified by enzyme cutting method. Then, pWH1-CXCR4 expression vector were transfected into salivary mucoepidermoid carcinoma Mc3 cells. At last, the expression of CXCR4 in Mc3 transfected with pWH1-CXCR4 expression vector was evaluated by RT-PCR and flow cytometry analysis. Result:Compared with Mc3 cells and Mc3 cells transfected with plasmid pWH1, the Mc3 cells transfected with pWH1-CXCR4 expression vector showed a lower mRNA and protein expression of CXCR4. Conclusion:The constructed CXCR4 shRNA expression vector can block the expression of CXCR4 in salivary mucoepidermoid carcinoma cells, and it may be helpful for further study on the significance of CXCR4 on the proliferation, metastasis and experimental treatment of salivary mucoepidermoid carcinoma.