Monoclonal antibody preparation and purification of a tumor cell collagenase-stimulatory factor.
Monoclonal antibody preparation and purification of a tumor cell collagenase-stimulatory factor.
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发表时间：
1989-06
期刊：
影响因子：
11.2
通讯作者：
Steven M. Ellis;Kazuki Nabeshima;Chitra Biswas
中科院分区：
文献类型：
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作者：
Steven M. Ellis;Kazuki Nabeshima;Chitra Biswas
来自几个来源的肿瘤细胞产生刺激成纤维细胞胶原酶产生的因子。已经产生了针对来自LX-1人肺癌细胞的肿瘤细胞胶原酶刺激因子的单克隆抗体，并且已经用于从LX-1细胞膜纯化该因子。这些纯化的制剂刺激成纤维细胞胶原酶的生产，80%的这些制剂含有一个单一的Mr约58，000的蛋白质可通过免疫印迹检测;其他20%含有一个额外的次要成分的分子量为35，000。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和放射自显影，在纯化因子的放射性标记制剂中也检测到分子量约为58，000的单一蛋白质。来自LX-1细胞的条件培养基含有几种分子量低于58，000的物质，其与膜衍生因子具有免疫交叉反应性。免疫荧光分析表明，肿瘤细胞胶原酶刺激因子分布在LX-1细胞的外表面，而成纤维细胞的细胞表面则不存在。这些和以前的结果表明，该因子是存在于肿瘤细胞表面上，被释放到条件培养基可能后，蛋白水解裂解，并出现在肿瘤侵袭过程中诱导宿主基质的胶原溶解的重要作用。
Tumor cells from several sources produce a factor(s) which stimulates fibroblast collagenase production. Monoclonal antibodies have been raised against the tumor cell collagenase-stimulatory factor from LX-1 human lung carcinoma cells and have been used for purification of the factor from LX-1 cell membranes. These purified preparations stimulated fibroblast collagenase production, and 80% of these preparations contained a single Mr approximately 58,000 protein detectable by immunoblotting; the other 20% contained an additional minor component with a molecular weight of 35,000. A single protein with a molecular weight of approximately 58,000 was also detected in radiolabeled preparations of the purified factor by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. Conditioned media from LX-1 cells contain several species with molecular weights lower than 58,000 which are immunologically cross-reactive with the membrane-derived factor. Immunofluorescence analysis indicates that the tumor cell collagenase-stimulatory factor is distributed on the outer surface of LX-1 cells and is absent from the cell surface of fibroblasts. These and previous results indicate that the factor is present on the tumor cell surface, is released into conditioned media possibly after proteolytic cleavage, and appears to have an important role in inducing collagenolysis of host stroma during tumor invasion.