Gualandris A, Lopez Conejo T, Giunciuglio D, Albini A, Sabini E, Rusnati M, Dell'Era P, Presta M
Department of Biomedical Sciences and Biotechnology, University of Brescia, Italy.
Microvasc Res. 1997 May;53(3):254-60. doi: 10.1006/mvre.1996.1998.
Fetal bovine aortic endothelial GM 7373 cells were transfected with a viral expression vector harboring the human urokinase-type plasminogen activator (uPA) gene. The stable transfectant clone uPA-R5 overexpressed and secreted human uPA as shown by Northern blot analysis, immunoprecipitation of metabolically labeled proteins, plasmin chromogenic assays, and SDS-PAGE zymography of cell extracts and conditioned media. The uPA-R5 cells were analyzed for their invasive capacity in vitro in the Matrigel chemoinvasion assay in the presence of serine- or metalloprotease inhibitors. uPA overexpression enhanced the invasive capacity of GM 7373 cells through a mechanism which differs from that mediated by metalloproteases. Endothelial cell uPA transfectants may represent an useful experimental model to investigate the role of uPA in angiogenesis and angioproliferative diseases.
用人尿激酶型纤溶酶原激活剂（uPA）基因的病毒表达载体转染胎牛主动脉内皮GM 7373细胞。Northern印迹分析、代谢标记蛋白的免疫沉淀、纤溶酶显色测定以及细胞提取物和条件培养基的SDS-PAGE酶谱分析表明，稳定转染克隆uPA-R5过表达并分泌人uPA。在存在丝氨酸或金属蛋白酶抑制剂的情况下，通过基质胶化学侵袭试验分析uPA-R5细胞的体外侵袭能力。uPA的过表达通过一种不同于金属蛋白酶介导的机制增强了GM 7373细胞的侵袭能力。内皮细胞uPA转染体可能是研究uPA在血管生成和血管增殖性疾病中作用的有用实验模型。
