Quaroni A, Isselbacher K J, Ruoslahti E
Proc Natl Acad Sci U S A. 1978 Nov;75(11):5548-52. doi: 10.1073/pnas.75.11.5548.
Synthesis of fibronectin in an epithelial cell line (IEC-6) established from rat small intestine was demonstrated by using immunofluorescence, radioimmunoassay, and collagen-binding. Internally labeled radioactive fibronectin isolated from the IEC-6 cells gave a single main band in sodium dodecyl sulfate/polyacrylamide gel electrophoresis under reducing conditions. Fibronectin isolated from rat plasma gave two closely spaced bands. The slower one had the same mobility as the epithelial cell fibronectin. The distribution of fibronectin in IEC-6 cells as detected by immunofluorescence was different from that described for fibroblasts and other cell types; fibronectin was present exclusively in regions of cell-to-cell contact. No fluorescence was detected on the surface membrane facing the culture medium or underneath the cells. This suggests that fibronectin may not be involved in the adhesion of the epithelial cells to the growth surface but could mediate cell-to-cell contacts. In microscopic sections of the small intestine, immunofluorescent staining with antifibronectin serum was strong in the basement membrane underlying the epithelial cells in the crypts. The in vitro synthesis of fibronectin by the crypt cells and its abundant presence in the basement membrane underlying the same cells in vivo suggests that fibronectin is a structural component of the basement membrane, and that it may be, at least in part, synthesized and deposited by the intestinal epithelium.
利用免疫荧光、放射免疫测定法和胶原结合法,证实了从大鼠小肠建立的上皮细胞系(IEC - 6)中纤连蛋白的合成。在还原条件下,从IEC - 6细胞中分离出的经内部标记的放射性纤连蛋白在十二烷基硫酸钠/聚丙烯酰胺凝胶电泳中呈现出一条单一的主带。从大鼠血浆中分离出的纤连蛋白呈现出两条紧密相邻的条带。较慢的那条带与上皮细胞纤连蛋白具有相同的迁移率。通过免疫荧光检测到的IEC - 6细胞中纤连蛋白的分布与成纤维细胞和其他细胞类型中所描述的不同;纤连蛋白仅存在于细胞间接触区域。在面向培养基的表面膜或细胞下方未检测到荧光。这表明纤连蛋白可能不参与上皮细胞与生长表面的黏附,但可能介导细胞间接触。在小肠的显微切片中,用抗纤连蛋白血清进行免疫荧光染色时,隐窝上皮细胞下方的基底膜中染色强烈。隐窝细胞在体外合成纤连蛋白以及其在体内相同细胞下方的基底膜中大量存在,表明纤连蛋白是基底膜的一种结构成分,并且它可能至少部分是由肠上皮合成和沉积的。