腺相关病毒载体介导的人内皮抑素的体外表达及对内皮细胞抑制作用的研究

Study on Adeno-associated Viral Vector-mediated Expression of Human Endostatin In Vitro and Inhibition of Endothelial Cell Proliferation

抗血管生成基因治疗是一有希望的抑制肿瘤生长及转移的方法。在实验中构建了含有人内皮抑素 (endo statin)基因的重组腺相关病毒载体rAAV -hE。所包装纯化的重组病毒滴度达 0 5× 10 12 v g /ml。rAAV -hE能有效感染培养细胞 ,EIA法检测培养液上清中内皮抑素浓度达 36 4 2ng/ml。rAAV介导所表达的内皮抑素对人脐静脉内皮细胞 (ECV - 30 4 )和牛毛细血管内皮细胞 (BCE)具有抗增殖抑制作用 ,抑制率分别为 6 8 1%和 4 1 6 %。此结果为进一步的动物实验奠定了基础 。

Antiangiogenesis gene therapy is a promising approach to inhibit tumor growth and metastases.We have constructed a recombinant adeno-associated viral vector,rAAV,which encodes a secretable form of human endostatin(hE).The titer of rAAV-hE was 0.5×10 12 v.g./ml.The rAAV-hE transduced the BHK-21 cells efficiently and EIA showed that the endostatin expression level was 36.42ng/ml.The supernatant from BHK-21 cells transduced by rAAV-hE showed inhibition activity on ECV-304 cell(HUVEC)and bovine capillary endothelial cell proliferation in vitro with the inhibitory rate of 68.1% and 41.6% respectively.These results offered a preliminary data for turther animal study and demonstrated the potential of recombinant adeno-associated viral vector-mediated antiangiogenic gene therapy in cancer therapy