ABSTRACT
Angiogenesis is an important process in physiology and disease pathogenesis and is controlled in a healthy body by a number of stimulatory and inhibitory factors. The aim of this study was to determine the effect of antisense transcript on the sense transcript of the endothelial growth factor [EGF] gene in bacterial system as an approach for the gene regulation in tumors. The hepatoma cell line [HepG2] was stimulated by PMA. VEGF mRNA was used for RT-PCR. VEGF cDNA was synthesised and cloned into T- vector pTZ57R, then sense fragment of VEGF subcloned into pACYC Duet-1 expression vector and antisense VEGF subcloned into pCDNAS expression vector. Recombinant plasmids were transforemed into BL2 1 bacterial cells. Expression of recombinant plasmid was analysed by western blot technique. The recombinant pCDNA3-VEGF [pYZantiVEGF] was successfully expressed in BL21 cells. Western blot analysis showed that the expression of VEGF decreased significantly in the cells transfected with VEGF antisense RNA compared with the pACYCDUET-1 - VEGF [pYZsenseVEGF] transfected and control. The expression of VEGF in BL21 cells was strong. In vitro, antisense of VEGF inhibited VEGF expression significantly in BL21 cells