胶原酶在肿瘤上皮细胞中的免疫定位

Immunolocalization of collagenase in neoplastic epithelial cells.

作者信息

Whitelock J M, O'Grady R L, Lyons J G

机构信息

Department of Cellular Pathology, University of Technology, Sydney, Gore Hill, NSW, Australia.

出版信息

Immunol Cell Biol. 1989 Dec;67 ( Pt 6):349-57. doi: 10.1038/icb.1989.51.

DOI:10.1038/icb.1989.51

PMID:2483153

Abstract

The distribution of interstitial collagenase in a rat mammary carcinoma model system has been studied by immunocytochemistry. Rabbit antibodies were raised against collagenase from neoplastic epithelial cells which were derived from an anaplastic, invasive, rat mammary carcinoma (BC1). Specificity of the antibodies was determined by Western blot analysis which showed reactivity with the inactive procollagenase from conditioned culture medium of BC1 cells as well as with purified, active BC1 collagenase. Anti-BC1 collagenase antibodies did not recognize BC1 collagenase entrapped by the inhibitor, rat alpha-2-macroglobulin (alpha 2M), or collagenase derived from TPA-stimulated human fibroblasts. Anti-human fibroblast collagenase antibodies did not recognize BC1 collagenase, suggesting that the human-mesenchymal and rat-epithelial enzymes are immunologically distinct molecules. Collagenase was immunolocalized intracellularly in BC1 cells cultured in the presence of monensin. Neither BC1 collagenase, alpha 2M nor enzyme-inhibitor complexes were demonstrated in or around invading tumours by immunostaining of tissue sections of rat mammary carcinomas.

摘要

通过免疫细胞化学方法研究了大鼠乳腺癌模型系统中间质胶原酶的分布情况。用源自一种间变性、侵袭性大鼠乳腺癌（BC1）的肿瘤上皮细胞中的胶原酶制备兔抗体。通过蛋白质印迹分析确定抗体的特异性，结果显示其与BC1细胞条件培养基中的无活性前胶原酶以及纯化的活性BC1胶原酶均有反应性。抗BC1胶原酶抗体不能识别被抑制剂大鼠α-2-巨球蛋白（α2M）捕获的BC1胶原酶，也不能识别源自佛波酯刺激的人成纤维细胞的胶原酶。抗人成纤维细胞胶原酶抗体不能识别BC1胶原酶，这表明人源间充质和大鼠上皮来源的酶在免疫上是不同的分子。在莫能菌素存在的情况下培养的BC1细胞内，胶原酶通过免疫定位法检测到。通过对大鼠乳腺癌组织切片进行免疫染色，在侵袭性肿瘤内或其周围均未检测到BC1胶原酶、α2M或酶-抑制剂复合物。