尿激酶及其受体对膀胱癌细胞系侵袭转移影响的体外研究

Effects of uPA and uPAR on the invasion and metastassis of bladder cancer cell lines in vitro

目的:探讨尿激酶型纤溶酶原激活物(uPA)及其受体(uPAR)与膀胱癌侵袭转移的关系。方法:对BIU-87、T24及EJ细胞系进行体外培养,夹心抗体ELISA法测定细胞培养上清及细胞溶解产物uPA含量,对培养细胞团块行uPAR免疫组化测定,并进行体外人工基底膜侵袭检测。结果:BIU-87、T24和EJ细胞培养上清uPA检测结果分别为(11.77±3.65)ng/ml、(8.70±2.45)ng/ml、(105.9±8.60)ng/ml,EJ细胞培养上清uPA明显高于BIU-87和T24细胞系(P<0.001)。细胞溶解产物uPA结果分别为(1.15±0.40)ng/ml、(0.78±0.34)ng/ml、(1.92±0.56)ng/ml,EJ细胞溶解产物uPA含量明显高于BIU-87、T24细胞溶解产物(P<0.02,P<0.01)。EJ细胞uPAR蛋白主要弥散于细胞质,着色强度与数量明显多于T24细胞(P<0.01),而BIU-87细胞基本无着色。EJ细胞具有浸润Matrigel的能力,BIU-87、T24无浸润Matrigel能力。结论:肿瘤细胞有uPA的分泌或来源并同时表达uPAR在体外即具备浸润能力,uPA系统在膀胱肿瘤的浸润转移过程中起重要作用。

Objective：To investigate the effects of urokinase-type plasminogen activator （uPA） and uPA receptor （uPAR） on the invasion and metastassis of bladder cancer cell lines in vitro. Methods： Quantitative analysis of uPA in culture medium and whole cell lysate of BIU 87, T24 and EJ human bladder carcinoma cell lines were assessed by enzyme linked immunosorbent assay （ELISA） ; Cell surface expression of the uPAR was observed by anti-uPAR monoclonal antibody using immunohistochemical technique; the invasive capacity of cells, with or without exogenous high molecular-weight urokinase, was examined in Matrigel invasion chambers. Results： The expressions of uPA in the three-bladder cancer cell lines are different. The concentration of uPA from EJ cells （105.9±8.60） ng/ml was the highest among three-bladder cancer cell lines and is significantly higher compared with that of BIU- 87（11.77±3.65）ng/ml,T24 （8.70±2.45）ng/ml cells （P〈0. 001,respectively）. In consistent, the cellular contain of uPA in EJ cell line （1.92±0. 56）ng/ml was significantly higher compared with that of BIU-87（1.15 ± 0. 40）ng/ml,P〈0. 02） and T24 （0.78±0.34）ng/ml, P〈0.01） cell line. In vitro EJ cell line, uPAR was mainly localized in the cytoplasm and the intensity of uPAR staining is significantly higher than that of T24 cell line （P〈 0.01） ; no expression was observed in BIU-87 cell line. EJ cells had the ability to invade Matriged whereas BIU-87 and T24 had not. Conclusions： This study demonstrates that bladder tumor cells express both uPA and uPAR. uPA and uPAR are required for bladder tumor cell invasion in vitro. Our data indicate that uPA and uPAR are both important for the clinical behavior of bladder neoplasms, which is possibly providing means for refined staging of muscle invasive tumors and target protein for novel therapies.