Varani J, Lovett E J, McCoy J P, Shibata S, Maddox D E, Goldstein I J, Wicha M
Am J Pathol. 1983 Apr;111(1):27-34.
High-metastatic murine fibrosarcoma cells readily attached to Type IV (basement membrane) collagen, whereas low-metastatic cells isolated from the same tumor did not. The addition of laminin--a glycoprotein that facilitates the adherence of epithelial cells to their basement membranes--enhanced the attachment of the low-metastatic cells, but not the high-metastatic cells. Using anti-laminin antibodies and a laminin-binding lectin as probes, the authors were able to identify by immunofluorescence a moiety associated with the high-metastatic cells, but not the low-metastatic cells, which cross-reacted with murine laminin purified from the EHS sarcoma. When extracts from the high-metastatic cells were separated by affinity chromatography, with the laminin-binding lectin as the affinity substrate, a substance was isolated that had an apparent molecular weight of 56,000 daltons. The affinity-purified material reacted strongly with anti-laminin antibodies by enzyme-linked immunosorbent assay.
高转移性小鼠纤维肉瘤细胞很容易附着于IV型(基底膜)胶原,而从同一肿瘤中分离出的低转移性细胞则不会。层粘连蛋白(一种促进上皮细胞附着于其基底膜的糖蛋白)的添加增强了低转移性细胞的附着,但对高转移性细胞没有作用。作者使用抗层粘连蛋白抗体和一种层粘连蛋白结合凝集素作为探针,通过免疫荧光法能够鉴定出一种与高转移性细胞相关而与低转移性细胞无关的部分,该部分与从EHS肉瘤中纯化的小鼠层粘连蛋白发生交叉反应。当用层粘连蛋白结合凝集素作为亲和底物,通过亲和层析分离高转移性细胞的提取物时,分离出一种表观分子量为56,000道尔顿的物质。通过酶联免疫吸附测定,亲和纯化的物质与抗层粘连蛋白抗体发生强烈反应。