Morioka S, Jensen P J, Lazarus G S
Exp Cell Res. 1985 Dec;161(2):364-72. doi: 10.1016/0014-4827(85)90093-x.
Using biochemical and immunocytochemical approaches, we have investigated the plasminogen activator (PA) of primary human epidermal cell cultures. A rabbit antibody raised against human urinary PA (urokinase) inhibited greater than or equal to 96% of the PA activity in the keratinocyte cultures. Immunoblot and double immunodiffusion analyses of keratinocyte PA with anti-urokinase antibody confirmed that epidermal PA was of the urokinase type. Immunocytochemical investigation of human keratinocyte cultures with anti-urokinase antibody revealed two characteristic staining patterns for PA. First, cells at the advancing edge of subconfluent colonies were cytoplasmically stained in a granular pattern. Similar staining was observed at the migrating edges of confluent epidermal cell cultures that had been wounded by cutting with a blade. This induction of PA staining was independent of cell division. Secondly, differentiated epidermal cells located on the surface of colonies were stained either at the plasma membrane or homogeneously throughout the cell. The highly differentiated, spontaneously shed cells were usually very heavily stained by anti-urokinase antibody. These immunocytochemical experiments suggest that PA expression is highly regulated in human epidermal cells. Specifically, PA expression appears to be related to cellular differentiation and to cell movement in expanding or wounded keratinocyte colonies.
我们采用生化和免疫细胞化学方法，对原代人表皮细胞培养物中的纤溶酶原激活剂（PA）进行了研究。一种针对人尿PA（尿激酶）产生的兔抗体，可抑制角质形成细胞培养物中96%及以上的PA活性。用抗尿激酶抗体对角质形成细胞PA进行免疫印迹和双向免疫扩散分析，证实表皮PA属于尿激酶型。用抗尿激酶抗体对人角质形成细胞培养物进行免疫细胞化学研究，发现PA有两种特征性染色模式。第一，亚汇合集落前沿的细胞胞质呈颗粒状染色。在用刀片切割造成损伤的汇合表皮细胞培养物的迁移边缘，也观察到类似的染色。这种PA染色的诱导与细胞分裂无关。第二，位于集落表面的分化表皮细胞，在质膜处或整个细胞内呈均匀染色。高度分化、自发脱落的细胞通常被抗尿激酶抗体强烈染色。这些免疫细胞化学实验表明，PA在人表皮细胞中的表达受到高度调控。具体而言，PA的表达似乎与细胞分化以及在扩展或受伤的角质形成细胞集落中的细胞运动有关。
