Raunio H, Konno R, Linnainmaa K, Wirth P J, Thorgeirsson S S
Laboratory of Experimental Carcinogenesis, National Cancer Institute, Bethesda, Maryland 20892.
Exp Cell Res. 1987 Dec;173(2):596-605. doi: 10.1016/0014-4827(87)90298-9.
The intercellular homotypic adhesive properties of 14 clones derived from a nontumorigenic rat liver epithelial cell line (LEC), derived from neonatal Fischer rats, were examined and compared to those of the hepatoma H4-II-E cell line. Each clone was assayed also for the degree of chromosomal aneuploidy and the ability to grow in soft agar. Over 100-fold differences in adhesive properties were observed among the clones, but no correlation was observed between the degree of aneuploidy in the clones and intercellular adhesive properties. The parent LEC cell line and the clones derived from it were unable to grow in soft agar. The H4-II-E cells showed negligible capacity to reaggregate after dissociation into single cells and these cells readily formed colonies in soft agar. Many of the LEC clones were similar to the H4-II-E cells in their adhesive properties, which suggests that reduced cell-to-cell adhesiveness per se is not a necessary prerequisite of epithelial cells to be able to grow independent of anchorage. Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) of concanavalin A (Con A)-binding glycoproteins in the "most adhesive" clone 67 and the "least adhesive" clone 201 showed markedly elevated amounts of acidic 105 and 67-kDa glycoproteins in clone 67. Proteins with similar migration patterns in 2D-PAGE have previously been reported to participate in specific homotypic intercellular adhesion of liver cells. The Con A-binding glycoprotein pattern in H4-II-E cells was markedly different from that of LEC cells with a set of six proteins missing and nine proteins appearing new in the H4-II-E cells. It is suggested that, in addition to identifying known epithelial cell polypeptides, systematic screening of cell surface-associated glycoproteins in normal and transformed epithelial cells in vitro and in vivo may lead to identification of novel polypeptides intimately associated with the transformed phenotype.
对源自新生Fischer大鼠的非致瘤性大鼠肝上皮细胞系(LEC)的14个克隆的细胞间同型黏附特性进行了检测,并与肝癌H4-II-E细胞系的黏附特性进行了比较。还对每个克隆的染色体非整倍体程度和在软琼脂中生长的能力进行了测定。各克隆间的黏附特性差异超过100倍,但克隆中的非整倍体程度与细胞间黏附特性之间未观察到相关性。亲本LEC细胞系及其衍生的克隆均不能在软琼脂中生长。H4-II-E细胞解离成单细胞后重新聚集的能力可忽略不计,并且这些细胞在软琼脂中很容易形成集落。许多LEC克隆在黏附特性上与H4-II-E细胞相似,这表明细胞间黏附性降低本身并非上皮细胞能够不依赖锚定而生长的必要先决条件。对“黏附性最强”的克隆67和“黏附性最弱”的克隆201中的伴刀豆球蛋白A(Con A)结合糖蛋白进行二维聚丙烯酰胺凝胶电泳(2D-PAGE)分析,结果显示克隆67中酸性105和67 kDa糖蛋白的含量明显升高。先前有报道称,在2D-PAGE中具有相似迁移模式的蛋白质参与肝细胞的特异性同型细胞间黏附。H4-II-E细胞中的Con A结合糖蛋白模式与LEC细胞明显不同,H4-II-E细胞中缺失一组六种蛋白质,同时出现九种新蛋白质。研究表明,除了鉴定已知的上皮细胞多肽外,在体外和体内对正常和转化上皮细胞中细胞表面相关糖蛋白进行系统筛选,可能会导致鉴定出与转化表型密切相关的新型多肽。