【摘要】 为研究腺相关病毒载体介导的内皮抑素(Endostatin)及K1-5蛋白抗肿瘤血管生成协同作用,实验中用含有Endostatin及K1-5基因的重组病毒rAAV-hE和rAAV-K5分别感染BHK-21细胞,并表达分泌出内皮抑素和K1-5蛋白,EIA法测定培养液上清中内皮抑素浓度达36.42ng/m l,免疫斑点印迹实验表明rAAV-K5可介导K1-5的体外表达。重组腺相关病毒(rAAV)介导所表达的内皮抑素和K1-5蛋白对血管内皮细胞增殖具有抑制作用。二者对人脐静脉内皮细胞(ECV-304)的抑制率分别为68.1%和63.1%,对牛毛细血管内皮细胞(BCE)的抑制率分别为41.6%和34.0%。同时二者还有协同效应,用rAAV-hE和rAAV-K5同时感染BHK-21细胞,表达产物对人脐静脉内皮细胞和牛毛细血管内皮细胞的抑制率分别为70.8%和43.8%。动物实验表明,重组病毒rAAV-hE和rAAV-K5转入荷有人肺肿瘤细胞的裸鼠,对肿瘤细胞生长具有抑制作用,同时二者具有协同抗肿瘤作用。更多还原

【Abstract】 To study on synergy of antiangiogenesis for tumor via adeno-associated viral vector-mediated gene transfer of endostatin and K1-5.Recombinant adeno-associated viral vectors rAAV-hE and rAAV-K5 transduced the BHK-21 cell and expressed endostatin and K1-5.Mean endostatin levels detecetd by EIA were 36.42ng/ml.Immuno-dot-blot assay showed that K1-5 gene was expressed in vitro.Conditioned supernatant from BHK-21 cell transduced with rAAV-hE or rAAV-K5 inhibited ECV-304 cell(HUVEC) and bovine capillary endothelial cell proliferation(BCE) in vitro.Inhibitory rates of HUVEC treated by two expressed proteins are 68.1% and 63.1%,and of BCE treated by two expressed protein are 41.6% and 34.0% respectively.In this paper we found that combination endostatin and K1-5 gene transfer induced synergistic inhibition on endotheial cells in vitro. This study showed that endostatin and K1-5 delivered by adeno-associated viral gene transfer could inhibit human lung carcinoma growth in nude mice.The study also demonstrated that combination endostatin and K1-5 gene transfer resulted in synergistic antitumor efficacy in human lung carcinoma in nude mice.更多还原