中文摘要：

目的研究核糖核酸酶抑制因子（RI）与整合素连接激酶（ILK）的相互作用以及对体外血管生成的影响。方法用LV5-RNH!homo、LV5NC、pCMV-3Xflag-ILK和pCMV-3×flag分别转染EJ细胞后，筛选稳定转染细胞系，分为EJ-RI、EJ-LV5、EJ-ILK、EJ-FLAG和EJ组；构建ILK与红色荧光蛋白融合表达载体YFP-ILK，与CFP-RI质粒共转染293细胞和EJ细胞，用荧光显微镜观察Rl和ILK蛋白在细胞内的共定位；构建真核表达质粒pCMV-3×flag-ILK，用GST蛋白沉降技术检测RI和ILK蛋白在体外的结合作用；用免疫共沉淀（co-IP）实验检测蛋向RI和ILK在真核细胞内的结合作用；用小管形成实验探索RI和ILK的相互作用对体外血管形成的影响。结果真核表达载体YFP-ILK和pCMV-3×flag-ILK构建成功；细胞内绿色荧光蛋白融合表达的RI和红色荧光蛋白融合表达的ILK在荧光显微镜下存在共定位现象；RI和ILK蛋白在原核细胞反应体系中存在相互作用；RI和ILK蛋白在真核细胞反应体系内也存在相互作用；RI和ILK的相互作用能抑制血管生成（P〈0．05）。结论RI和ILK蛋白在原核细胞防御体系和真核细胞反应体系中均存在相互结合作用，并对体外血管生成有抑制作用。

英文摘要：

Objective To research the interaction between ribonuclease inhibitor（RI） and inte-grin-linked kinase （ILK） and its effect on angiogenesis in vitro. Methods To construct YFP-ILK and pCMV-3 × flag-ILK plasmid. Purchase lentiviral expres-sion vector LVS-RNH1 homo and LVSNC. Stably transfected vectors into EJ cells repec- tively, selected cells were named as EJ-RI, E J-ILK, EJ-LV5, E J-FLAG and EJ. Fluorescence microscope analysis was used to observe the Co-location between RI and ILK in 293 cells and EJ cells. The interaction between RI and ILK in 293 and EJ cells was identified by glutathione-S-transferase （GST） pull-down assay and co-immunoprecipi- ration assay. Endothelial tube formation assay was used to assess angiogenesis in vitro. Results The expressing pl- amid YFP-ILK and pCMV-3 × flag-ILK were verified by restriction enzyme digestion and DNA sequencing. We found that RI was co-localized with ILK in 293 and EJ cell. Glutathione-S-transferase （GST） pull-down andco-immunoprecipitation results showed that the interaction between RI and ILK occurred in prokaryotic cells and mammalian cells as well. RI can inhibit angiogenesis in vitro by interacting with ILK. Conclusions The interaction between RI and ILK occurs in prokatyotic cells and mammalian cells and inhibits angiogenesis in vitro.