Ohshima S, Ishimaru Y, Honda M, Ohkawara S, Ogawa M
Second Department of Surgery, Kumamoto University School of Medicine, Japan.
Cell Tissue Res. 1993 Aug;273(2):363-70. doi: 10.1007/BF00312839.
A cell surface-associated adhesive factor (AF) separated from differentiated rat ascites hepatoma AH136B cells (forming cell islands in vivo) has been highly purified by chromatography. AF is assumed to mediate the cell-cell adhesion essential to island formation of the hepatoma cells. A substance, immunologically cross-reactive with AF, is present in the ascites fluid or culture medium of the AH136B cells. Because the substance is almost identical to AF in molecular weight and aggregation-promoting activity, it has been concluded that AF is released into the ascites fluid where it is concentrated. Monoclonal antibodies have been raised against AF purified from ascites fluid of AH136B cells. We have obtained a monoclonal antibody, coded MoAF-6D6, that strongly abolishes the aggregation-promoting activity of AF. When AH136B cell islands are incubated in the presence of Fab fragments of MoAF-6D6, cell detachment from the islands is evident within 24 h. Cell islands following 36-h culture show a distinct dissociation and islands completely lose their organization 48 h after culture. The dissociating effect of MoAF-6D6 is neutralized by the addition of AF. These results suggest that AF plays a significant role in the maintenance of cell islands.
从分化的大鼠腹水肝癌AH136B细胞(在体内形成细胞岛)中分离出的一种细胞表面相关粘附因子(AF)已通过色谱法高度纯化。AF被认为介导了肝癌细胞形成细胞岛所必需的细胞间粘附。在AH136B细胞的腹水或培养基中存在一种与AF免疫交叉反应的物质。由于该物质在分子量和促进聚集活性方面与AF几乎相同,因此得出结论,AF被释放到腹水中并在其中浓缩。已经制备了针对从AH136B细胞腹水中纯化的AF的单克隆抗体。我们获得了一种编码为MoAF-6D6的单克隆抗体,它能强烈消除AF的促进聚集活性。当在MoAF-6D6的Fab片段存在下孵育AH136B细胞岛时,24小时内细胞从细胞岛脱离明显可见。培养36小时后的细胞岛显示出明显的解离,培养48小时后细胞岛完全失去其组织结构。加入AF可中和MoAF-6D6的解离作用。这些结果表明,AF在维持细胞岛方面起着重要作用。