Volume 19, Number 3 (October 2012)                   J Birjand Univ Med Sci. 2012, 19(3): 255-265 | Back to browse issues page


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Zarei Mahmud abadi A, Masoomi Karimi M, Bahabadi M, Kamali M, Kushki R, Khalili F, et al . Preventing the expression of VEGFR-1 in culture medium using specific SiRNA - as a potential therapeutic method in eye neovascularization. J Birjand Univ Med Sci.. 2012; 19 (3) :255-265
URL: http://journal.bums.ac.ir/article-1-1061-en.html

1- PhD of Clinical Biochemistry Biochemistry Department& Chemical Damage Research Center, Baghiyatallah University of Medical Science, Tehran, Iran.
2- MSc of Immunology , Immunology Department, torbateheydariyeh university of medical science. Torbateheydariyeh, Iran
3- MSc of Clinical Biochemistry, Biochemistry Department, Baghiyatallah University of Medical Science, Tehran, Iran
4- PhD of Biotechnology. Nano-biotechnology Research Center, Baghiyatallah University of Medical Science, Tehran, Iran
5- MSc of Cellular and Molecular Biology, Nano-biotechnology Research Center, Baghiyatallah University of Medical Science, Tehran, Iran.
6- MD of Retina, Department of Ophthalmology, Baghiyatallah University of Medical Science, Tehran, Iran.
7- MS of cellular and molecular biology, Nano-biotechnology Research Center, Baghiyatallah University of Medical Science, Tehran, Iran
8- Corresponding author, PhD Student of Clinical biochemistry, biochemistry department, torbateheydariyeh university of medical science. Torbateheydariyeh, Iran
9- MSc of Nano-biotechnology Nano-biotechnology Research Center, Baghiyatallah University of Medical Science, Tehran, Iran.
Abstract:   (11002 Views)
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<0.05 was taken as the significant level. Results: The fluorescence emission from defined siRNA decreased compared to control group. SDS pages and blots from vector cloned cells exposed to both siRNA showed reduced protein expression The outcome of applying two siRNA indicates gene expression in the form of transcription and translation, compared to the control group (P<0.05). Conclusion: Specifically designed siRNA against VEGFR1, through lipofectamin, was appropriately transferred into cell and significantly prevented from the receptor expression. In fact, by blocking angiogenesis signaling route, it was able to prevent neovascurization. Thus, this can be made use of as an appropriate factor in preventing or decreasing neovascularization in the eye.
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Type of Study: Original Article | Subject: Biochemistry
Received: 2012/03/4 | Accepted: 2012/10/31 | Published: 2016/03/10

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