Angiostatin基因转染对膀胱癌BIU-87细胞系生物学行为的影响

目的 探讨外源性血管抑素(angiostatin)基因在人膀胱癌BIU-87细胞中的表达及其意义。方法 将人全长angiostatin基因经脂质体包裹转染膀胱癌BIU-87细胞，行G418筛选获得转基因瘤细胞克隆，比较转基因和未转基因的瘤细胞生长特性。采用免疫荧光、免疫组化及Western blot等方法检测瘤细胞angiostatin蛋白的表达，并应用鸡胚尿囊膜血管生成实验及脐静脉内皮细胞增殖实验检测其活性。结果 外源性angiostatin基因的表达对BIU-87瘤细胞的生长并无影响。转染细胞在体外明显抑制脐静脉内皮细胞的增殖并对鸡胚尿囊膜的血管生成具有明显的抑制作用。结论Angiostatin能特异性地抑制血管内皮细胞增殖，进而抑制血管生成。

Effects of angiostatin gene transfection on biological behaviors of bladder carcinoma BIU-87 cell line

Objective To explore the expression and effects of extragenous angiostatin gene in human bladder carcinoma BIU-87 cell line. Methods A full human angiostatin gene was cloned into expression vector pcDNA3.1(+) and transfected into human bladder carcinoma BIU-87 cells by lipofectamine, and the positive clone was screened by G418. The growth characteristics of BIU-87 cells before and after transfection were compared.The expressions of angiostatin protein in BIU-87 cells were examined by immunofluorescence assay, immunohistochemistry and Western blotting, and the activities of angiostatin protein were examined by endothelial cell inhibition assay and the chicken-embryo chorioallantoic membrane assay. Results The expression of extragenous angiostatin gene had no inhibitory effect on the growth of BIU-87. The transfected cells could inhibit the umbilical vein endothelial cell growth in vitro and chicken-embryo choriallantoic membrane angiogenesis in vivo. Conclusion The results indicated that the angiostatin gene clones could be transfected into BIU-87 bladder cancer cells, and angiostatin could inhibit endothelial cell proliferation and neovascularization.