Inhibition of VEGFR-1 gene expression in U937 cell line using Lentivirus-mediated shRNA

Objective To construct the shRNA expression vector targeting human vascular endothelial growth factor receptor 1(VEGFR-1) gene and to detect its silencing effects on human monocytic leukemia cell line U937.Methods Three pairs of target segments were synthesized and cloned into pRNAT-U6.2 Lentiviral vector respectively and the vectors were identified by PCR and DNA sequencing.Then the expression vectors were transferred into U937 cell line to produce packaged lentivirus.After being infected with the packaged lentivirus,the expression of VEGFR-1 gene of U937 cell line at mRNA and protein level was detected by Real-Time PCR and Western blotting. Results PCR and DNA sequencing showed that the target segments were cloned into pRNAT-U6.2 lentiviral vector respectively.The expression rate of VEGFR-1 gene in U937 cells in the RNA interference group had dropped by 75.98%as compared to that of the control by Real-Time PCR. Western blot also showed decreased expression of the VEGFR-1 protein in the RNA interference group.Conclusion The shRNA expression vector effectively inhibits the expression of VEGFR-1 gene in U937 cell line on mRNA and protein levels.