The siRNA-Mediated Down-Regulation of Vascular Endothelial Growth Factor Receptor1.
BACKGROUND
Angiogenesis is an important biological process involved in the proliferation of endothelial cells, tumor growth and metastasis. Vascular endothelial growth factor (VEGF) is considered as a prominent regulator of angiogenesis which exerts the aforementioned effect(s) through its respective receptors (VEGFR1 and VEGFR2). VEGF receptors are targeted as a therapeutic candidate for cancer growth inhibition. RNAi as a new and promising strategy has provided a useful means to specifically suppress gene expression in cancer cells.


OBJECTIVE
The current study aimed to down-regulate expression of the VEGFR1 using siRNA.


METHODS
This experimental study designed specific siRNAs against VEGFR1. Total RNA was extracted from human umbilical vain endothelial cell (HUVEC) and subsequently cDNA was synthetized. PCR was performed using specific primers to amplify the target gene. After double digestion and purification, the gene was cloned into pEFGP-N1 expression vector. Then, AGS cells were transfected with recombinant pEGFP-N1 using lipofectamin. The gene expression and down-regulation were evaluated by fluorescence scanning, reverse transcription PCR (RT-PCR) and Western blot techniques.


RESULTS
Fluorescent scanning, RT-PCR (27.68%) and western blot analysis (31.06%) showed that the expression of VEGFR1 was suppressed effectively.


CONCLUSIONS
The results of the current study showed that specifically designed siRNA can be considered as an appropriate strategy to suppress gene expression and might be a promising tool to prevent angiogenesis.
Biochemistry Department, School of Medicine, Shahroud University of Medical Sciences, Shahroud, IR Iran.
Moslem
M
Jafari Sani
Biotechnology Department, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, IR Iran.
Foad
F
Yazdi
Immunology Department, School of Medicine, Torbat Heydariyeh University of Medical Sciences, Torbat Heydariyeh, IR Iran.
Masoomeh
M
Masoomi Karimi
Department of Molecular Medicine, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, IR Iran.
Javad
J
Alizadeh
Biotechnology Department, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, IR Iran.
Majid
M
Rahmati
Biochemical Department, Chemical Injuries Research Center, Baqiyatallah University of Medical Sciences, Tehran, IR Iran.
Ali
A
Zarei Mahmudabadi
Angiogenesis; Gastric Cancer; Small Interfering RNA; Vascular Endothelial Growth Factor Receptor