Taylor-Papadimitriou J, Burchell J, Hurst J
Cancer Res. 1981 Jun;41(6):2491-500.
The production and retention of fibronectin by primary cultures of cells derived from the human breast has been analyzed. Two examples of each of the following cell types were examined: (a) normal epithelium from milk; (b) metastatic breast cancer cells in pleural effusions; (c) fibroblasts; (d) tissue macrophages of milk. Cell-associated fibronectin could be detected by indirect immunofluorescent staining on normal and malignant mammary epithelium and on mammary fibroblasts, but not on milk macrophages. Immune precipitation followed by gel electrophoresis of 35S-labeled cell lysates and conditioned medium confirmed that fibronectin was indeed synthesized by both types of epithelial cells and by fibroblasts, but not by macrophages, and that much of the protein was released into the medium. Quantitative analysis with radioimmune assay of the fibronectin on cells and in media showed that both normal and malignant epithelial cells synthesized levels of protein comparable to that produced by fibroblasts, but only a small fraction (less than 10%) of the material synthesized was retained by the cells. Growth on collagen-coated plastic increased the percentage of fibronectin retained by normal and malignant epithelium but did not affect retention by fibroblasts.
对源自人乳腺的细胞原代培养物中纤连蛋白的产生和保留情况进行了分析。对以下每种细胞类型各选取两个实例进行了检测:(a) 乳汁中的正常上皮细胞;(b) 胸腔积液中的转移性乳腺癌细胞;(c) 成纤维细胞;(d) 乳汁中的组织巨噬细胞。通过间接免疫荧光染色可在正常和恶性乳腺上皮细胞以及乳腺成纤维细胞上检测到细胞相关纤连蛋白,但在乳汁巨噬细胞上未检测到。对35S标记的细胞裂解物和条件培养基进行免疫沉淀后再进行凝胶电泳,证实纤连蛋白确实由两种上皮细胞和成纤维细胞合成,而非巨噬细胞合成,并且大部分蛋白质释放到了培养基中。用放射免疫分析法对细胞和培养基中的纤连蛋白进行定量分析表明,正常和恶性上皮细胞合成的蛋白质水平与成纤维细胞产生的相当,但细胞仅保留了合成物质的一小部分(不到10%)。在胶原包被的塑料上生长增加了正常和恶性上皮细胞保留的纤连蛋白百分比,但不影响成纤维细胞的保留情况。