Establishment Of VEGF-transfected Cell Line And Study On Its Radiobiological Effects

Object: It has been reported that VEGF can selectively stimulate endothelial cells proliferate and prevent them apoptosis. However, the role of VEGF on radiation-induced vascular endothelial cell injure remains unknown. In this study, we demonstrated the biological characters of vascular endothelial cells and identify radiobiological characters of VEGF-transfected cell line, which could get a new insight into the mechanism and value to find potential therapeutic targets in radiation bum.Methods: The proliferation and chromosome damage of vascular endothelial cells treated by 7 ray was examed according to ³H-TdR incorporation and micronucleus. The expression of VEGF mRNA and protein after irradiation was detected by RT-PCR and western blot. Expression vector of VEGF₁₆₅ sense and antisense gene was transfected into vascular endothelial cells. Stable cell lines were established and its radiobiological effects were evaluated using MTT assay, micronucleus and change of mitochondrial membrane potential.Results: ³H-TdR incorporation decreased after 7 ray irradiation, whlie micronucleus and levels of VEGF mRNA and protein expression increased, which all appeared in dose-dependent manner. The level of VEGF mRNA was elevated after 6h when 8Gy irradiation was processed and maintained to 72h. Sense and anti-sense expression vector were transfected into vascular endothelial cells and stable cell lines were established. Cell line transfected sense expression vector of VEGF₁₆₅ gene showed reduced inhibition of prolifetation, MNF, MNCF and apoptosis compared to that of control vector, while opposite results were obtained in anti-sense transfected cell line.Conclusion: VEGF has a protective effect on the vascular endothelial cells to 7 ray irradiation, which may illustrated a new therapy in irradiation injure.