ROLE OF LAMININ IN THE ATTACHMENT OF PAM212 (EPITHELIAL) CELLS TO BASEMENT-MEMBRANE COLLAGEN
ROLE OF LAMININ IN THE ATTACHMENT OF PAM212 (EPITHELIAL) CELLS TO BASEMENT-MEMBRANE COLLAGEN
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DOI：
10.1016/0092-8674(80)90548-6
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发表时间：
1980-01-01
期刊：
影响因子：
64.5
通讯作者：
MARTIN, GR
中科院分区：
文献类型：
--
作者：
TERRANOVA, VP;ROHRBACH, DH;MARTIN, GR
对小鼠上皮细胞系PAM-212在不同胶原基质上的黏附测定表明，这些细胞优先黏附在基底膜IV型胶原上。血清和纤维连接蛋白都不能刺激这些细胞(不同于成纤维细胞)与IV型胶原的附着。用放线菌酮预先孵育PAM 212细胞可防止附着。这些细胞不通过血清中存在的大分子进行附着，而是合成一种附着因子。EHS肿瘤的提取物产生了含有基底膜成分的细胞外基质，测试了它们促进与环己亚胺处理的PAM 212细胞附着的能力。在放线菌胺存在的情况下，肿瘤的生理盐水提取物可刺激PAM212细胞与IV型胶原的附着。层粘连蛋白是一种高相对分子质量的基底膜糖蛋白成分，是从盐提取物中提纯的活性物种，其浓度低至1-5微克/毫升。当层粘连蛋白预先孵育在涂有I、II、III、IV或V型胶原的平板上并随后清洗时，只有IV型胶原涂层的平板上才能看到高水平的附着。针对层粘连蛋白的亲和纯化抗体抑制PAM 212细胞与IV型胶原底物的附着。层粘连蛋白似乎是一种特殊的上皮细胞附着蛋白，因为它不能刺激成纤维细胞与I型或IV型胶原基质的附着。显然，层粘连蛋白是由这些上皮细胞产生并利用来附着到基底膜胶原上的。
Measurements of the attachment of the PAM 212 line of mouse epithelial cells to various collagen substrates show that these cells adhere preferentially to type IV, basement membrane collagen. Neither serum nor fibronectin stimulated the attachment of these cells (unlike fibroblasts) to type IV collagen. Preincubation of the PAM 212 cells with cycloheximide prevented attachment. These cells do not attach by means of a macromolecule present in serum but instead synthesize an attachment factor. Extracts of the EHS tumor, which produces an extracellular matrix containing basement membrane components, were tested for their ability to promote attachment to cycloheximide-treated PAM 212 cells. Saline extracts of the tumor stimulated the attachment of the PAM 212 cells to type IV collagen in the presence of cycloheximide. Laminin, a high MW glycoprotein constituent of basement membrane, was purified from the salt extract and was the active species at concentrations as low as 1-5 .mu.g/ml. When laminin was preincubated on plates coated with either type I, II, III, IV or V collagen and the plates subsequently washed, high levels of attachment were seen only on type IV collagen-coated plates. Affinity purified antibody directed against laminin inhibited the attachment of PAM 212 cells to a type IV collagen substrate. Laminin appears to be a specific attachment protein for epithelial cells since it did not stimulate the attachment of fibroblasts to type I or to type IV collagen substrates. Apparently, laminin is produced and used by these epithelial cells to attach to basement membrane collagen.