重组腺病毒介导VEGF_(121) cDNA体外转移与表达

Recombinant Adenovirus-mediated VEGF_(121) cDNA Gene Transfer and Expression In vitro

构建携带VEGF12 1cDNA重组复制缺陷型腺病毒表达载体 ,制备重组腺病毒 ,该腺病毒能介导VEGF12 1cDNA基因促进人脐静脉血管内皮细胞增殖 ,促进毛细血管管腔样结构形成 .ELISA检测表明VEGF12 1cDNA基因表达产物分泌至培液上清 ,并显示强烈血管通透作用 .为进一步利用重组腺病毒介导VEGF12

The replication-incompetent adenoviral vector containing the cDNA of VEGF(121) was constructed. After transfection, individual viral plaques were isolated and amplified in HEK293 cells. Confirmed by PCR, both Adeno-X-VEGF(121) and Adeno-X-LacZ were propagated in HEK293 cells and were purified by CsCl density gradient centrifugation. Adenovirus-mediated VEGF(121) gene transfer promotes ECV304 proliferation and formation of capillary-like structures in vitro. The expression of VEGF(121) by adenovirus-infected ECV304 was quantified by enzymelinked immunosorbent assay. Peak VEGF(121) production was achived at 7similar to10 days after infection. The conditioned media from ECV304 infected with Adeno-X-VEGF(121) markedly enhanced vascular permeability. These data may support that adenovirus-mediated VEGF(121) cDNA gene transfer could provide a useful strategy for efficient delivery of VEGF(121) in the treatment of ischemic diseases.