Preventing the expression of VEGFR-1 in culture medium using specific SiRNA - as a potential therapeutic method in eye neovascularization

Background and Aim: Angiogenesis is one of important biological processes any disruption in which leads to disease. The main signaling factor in this process is VEGF which acts through its receptors. The present study was done in order to inhibit the expression of receptor type1of this factor (VEGFR-1) using specific siRNA in the culture medium to use its inhibitory effect on neovascularization in the eye. Materials and Methods: In this experimental study first, using target gene sequences, sequences of the specific siRNA were designed against them blasted and manufactured. On the other hand, cDNA of HUVEC cell was synthesized and PCR, with specific primers for target gene, was reproduced as necessary. Then, PEFGP-N1 expression vectors were cloned and confirmed. Then, the obtained plasmid vector was transferred to Hela cells lacking target expressive genes through lipofectamin. GFR expression rate in the initial vector and in the cloned one, both in presence and in absence of specific VEGFR1 siRNA, was assessed. Evaluation of gene inhibition was carried out through decreasing of green fluorescence from GFR, Western blot and RT-PCR. Results were analyzed using T-test and P