Inhibition of VEGF gene expression in Hela cell by short interfering RNAs
- VernacularTitle:siRNA对Hela细胞表达VEGF的干涉作用
- Author:
Bin FAN
;
Guangyu LI
;
Yan TAN
- Publication Type:Journal Article
- Keywords:
RNA interference;
T7 RNA ploymerase;
VEGF;
Hela cells
- From:
Chinese Journal of Immunology
2000;0(08):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the interfering effect of siRNA on the expression of VEGF in Hela cells.Methods:Three pairs of siRNA were designed according to the 1-5 extrons sequence of VEGF and synthesized by T7 RNA ploymerase transcription in vitro. To evaluate the inhibition activity of siRNA, Hela cells were transfected via siPORT Lipid. The interfering effect of siRNAs in hRPE cells was examined by semi-quantitative RT-PCR and immunofluorescence technique.Results:The results showed that the three pairs of siRNA could effectively inhibit gene expression of VEGF in Hela cells with the rates of 88.7%, 94.2% and 80.3%. But the 1-base mismatched siRNA and ssRNA(+) didn′t show significant interfering effect. Furthermore, it was found that siRNAs had a dose dependent inhibitory effect(5-10 pmol).Conclusion:The siRNAs synthesized by T7 RNA ploymerase in vitro could effectively and specifically interfere the expression of VEGF in Hela cells, providing a novel approach for gene therapy of tumor.
