Christensen L, Wiborg Simonsen A C, Heegaard C W, Moestrup S K, Andersen J A, Andreasen P A
Department of Pathology, Glostrup Hospital, Denmark.
Int J Cancer. 1996 May 16;66(4):441-52. doi: 10.1002/(SICI)1097-0215(19960516)66:4<441::AID-IJC6>3.0.CO;2-W.
We have investigated the localization of urokinase-type plasminogen activator (u-PA), type-1 plasminogen-activator inhibitor (PAI-1), u-PA receptor (u-PAR) and alpha(2)-macroglobulin- receptor/low-density-lipoprotein-receptor-related protein (alpha(2)MR/LRP) in human breast tumors by immunohistochemical methods. Frozen sections of 133 primary breast carcinomas, 6 ductal carcinomas in situ and 33 lymph-node metastases were stained with monoclonal antibodies. Formalin-fixed sections of 15 primary tumors and 2 lymph-node metastases were stained with polyclonal antibodies. In primary tumors, u-PA and PAI-1 immunoreactivities were intense in macrophages and mast cells, and moderate in benign and malignant epithelial cells as well as in myofibroblasts and endothelial cells. A sub-group of poorly differentiated tumors showed particularly strong staining of stromal fibroblasts. u-PA immunoreactivity was also present in lymphocytes. alpha(2)MR/LRP and u-PAR immunoreactivities were intense in macrophages, but apart from these cells, alpha(2)MR/LRP was found only in fibroblasts, and u-PAR only in tumor cells located peripherally in tumor-cell clusters and glands and some myofibroblasts in the adjacent stroma. Lymph-node metastases showed staining for u-PA and PAI-1 both of cancer cells and of stromal fibroblasts, also staining for u-PA of lymphocytes. Similarly to some of the poorly differentiated primary tumors, approximately half of the metastases showed very strong staining of stromal fibroblasts, and extracts of these metastases had higher u-PA and PAI-1 levels, as determined by ELISA, than extracts of metastases without this staining pattern. alpha(2)MR/LRP was present only in fibroblasts and u-PAR only in some tumor cells. The presence of u-PA, PAI-1, alpha(2)MR/LRP and u-PAR was controlled biochemically by immunoblotting analyses, ligand-blotting analyses, and direct and reverse zymography. The spatial distribution and the variation in concentration of the various components of the plasminogen-activation system point to a complex, multifunctional role for the 4 proteins in and/or during the development and spread of breast cancer.
我们采用免疫组化方法研究了尿激酶型纤溶酶原激活剂（u-PA）、1型纤溶酶原激活剂抑制剂（PAI-1）、u-PA受体（u-PAR）以及α2-巨球蛋白受体/低密度脂蛋白受体相关蛋白（α2MR/LRP）在人乳腺肿瘤中的定位。用单克隆抗体对133例原发性乳腺癌、6例原位导管癌和33例淋巴结转移灶的冰冻切片进行染色。用多克隆抗体对15例原发性肿瘤和2例淋巴结转移灶的福尔马林固定切片进行染色。在原发性肿瘤中，u-PA和PAI-1免疫反应性在巨噬细胞和肥大细胞中强烈，在良性和恶性上皮细胞以及肌成纤维细胞和内皮细胞中中等。一组低分化肿瘤的基质成纤维细胞染色特别强。u-PA免疫反应性也存在于淋巴细胞中。α2MR/LRP和u-PAR免疫反应性在巨噬细胞中强烈，但除这些细胞外，α2MR/LRP仅在成纤维细胞中发现，u-PAR仅在肿瘤细胞簇和腺体周边的肿瘤细胞以及相邻基质中的一些肌成纤维细胞中发现。淋巴结转移灶中癌细胞和基质成纤维细胞均显示u-PA和PAI-1染色，淋巴细胞也显示u-PA染色。与一些低分化原发性肿瘤类似，约一半的转移灶基质成纤维细胞染色非常强，通过ELISA测定，这些转移灶的提取物中u-PA和PAI-1水平高于无此染色模式的转移灶提取物。α2MR/LRP仅存在于成纤维细胞中，u-PAR仅存在于一些肿瘤细胞中。通过免疫印迹分析、配体印迹分析以及直接和反向酶谱分析对u-PA、PAI-1、α2MR/LRP和u-PAR的存在进行了生化检测。纤溶酶原激活系统各组分的空间分布和浓度变化表明这4种蛋白在乳腺癌发生发展和扩散过程中具有复杂的多功能作用。
