【Objective】 To construct human vascular endothelial growth factor-165 (hVEGF165) gene lentivirus expression vector pCDH-CMV-MCS-EF1-copGFP-hVEGF165

and detect its expression after transfected into HT22 cells and further observe its effect of anti-apopotosis induced by irradiation. 【Methods】 hVEGF165 gene cDNA was amplicated by polymerase chain reaction (PCR) and cloned into the pCDHCMVMCSEF vector. Reconstructed pCDH-CMV-MCS-EF1-hVEGF165 plasmid was identified by electromigratory analysis

colony PCR

and sequencing analysis. 293T cells were transfected with recombinate vector using the way of calcium phosphate cotransfection to get lentiviral particles. Lentiviral particles were then transfect them into HT22 cells. Real-time RT-PCR and Western blot were used to detect the expression of VEGF165 gene in HT22 cells. Furthermore

0 Gy

5 Gy

and 10 Gy X-ray irradiation were given to HT22 cells

empty vector transfected HT22 cells and hVEGF165 gene transfected HT22 cells

respectively. Flow cytometry was then used to compare different rates of apoptosis.【Results】 hVEGF165 gene fragment was constructed into the pCDH-CMV-MCS-EF1-copGFP vector successfully. Enzyme digestion got correct length of hVEGF165 gene

DNA sequencing analysis confirmed that hVEGF165 gene sequence was exactly the same with that reported by GenBank. There were significant increased expression of hVEGF165 mRNA and protein in the HT22 cells after transfected. Compared with the control group

hVEGF165 could reduce apoptotic rate after cell radiation. 【Conclusions】 pCDH-CMV-MCS-EF1-copGFP-hVEGF165 lentivirus vector has been successfully constructed. High level of hVEGF165 mRNA and protein expression could be obtained in HT22 cells after transfected with pCDH-CMV-MCS-EF1-copGFP-hVEGF165. hVEGF165 gene can protect the cells from irradiation induced apoptosis.