Chinese Medical E-ournals Database

Chinese Journal of Obstetrics & Gynecology and Pediatrics(Electronic Edition) ›› 2007, Vol. 03 ›› Issue (03): 129 -f3. doi: 10.3877/cma.j.issn.1673-5250.2007.03.103

Original Article

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

Hai-xia GUO, Hong-man XUE, Yang LI, Zhe MENG, Huan CHEN, Wen-yi LI   

  1. The Department of Pediatrics, the Second Affiliated Hospital, Sun Yat-Sen University, Guangzhou, 510120, China
  • Published:2007-06-01
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.

图1 VEGFR-2 shRNA表达载体对HL60 VEGFR-2表达影响的Western blot分析
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