Design of human vascular endothelial growth factor receptor 2 siRNA and construction of its expression vector

Objective To search and investigate human vascular endothelial growth factor receptor 2 (VEGFR-2) small interference RNA (siRNA) with strong interfering effect to suppress leukemia cell. Methods Designed human VEGFR-2 siRNA and screened the most efficient with MTT method after transfecting HL60 cells ; synthesized shRNA and constructed VEGFR-2 shRNA expression vector ; compared the interfering effects after transfecting HL60 cell with VEGFR-2 shRNA expression vector. Results The most efficient siRNA was screened ; VEGFR-2 shRNA was cloned correctly into pENTR TM/U6 and constructed VEGFR-2 shRNA expression vector successfully; transfecting HL60 cell transiently with it, cell inhibitory rate was as high as 84. 9%, P<0. 01; expression of VEGFR-2 protein decreased significantly. Conclusion SiRNA target sequence with relatively good inhibitory effect should follow necessary and effective screening principals. VEGFR-2 shRNA expression vector can clone shRNA simply and quickly in cells and has high inhibitory rate of VEGFR-2 expression. Key words: small interference RNA (siRNA); vascular endothelial growth factor receptor 2(VEGFR-2); HL60; expression vector