Hart D A, Smith R
Cancer Lett. 1987 Apr;35(1):27-38. doi: 10.1016/0304-3835(87)90053-x.
Analysis of conditioned medium from three sublines of the B16 melanoma [F1 (parental), BL6 (invasive), F10 (metastatic)] by SDS-PAGE and zymography revealed the presence of plasminogen activator activity at 60,000 daltons. The relative activity was F10 greater than F1 greater than or equal to BL6. Treatment of the cells with the tumor promoter, phorbol myristate acetate (PMA) led to increased secretion of PA by F10 cells and a lesser increase in secretion by F1 cells and BL6 cells. In addition, a second plasminogen activator activity at 45,000 daltons was detected in conditioned medium from PMA treated F10 cells. Conditioned medium from F10 and F1 cells was also shown to contain a 33,000 dalton plasminogen activator binding protein. Upon PMA treatment the concentration of the binding protein increased in medium from F10 cells but not in similarly treated F1 cells. The binding protein, very likely a plasminogen activator inhibitor, was nearly undetectable in conditioned medium from control and PMA-treated BL6 cells. Therefore, the three sublines, which differ in in vivo phenotypic characteristics, also differ in their in vitro regulation of proteinase and proteinase inhibitor synthesis.
通过SDS-聚丙烯酰胺凝胶电泳（SDS-PAGE）和酶谱分析对B16黑色素瘤的三个亚系[F1（亲本）、BL6（侵袭性）、F10（转移性）]的条件培养基进行分析，结果显示在60,000道尔顿处存在纤溶酶原激活物活性。相对活性为F10大于F1大于或等于BL6。用肿瘤启动子佛波醇肉豆蔻酸酯乙酸酯（PMA）处理细胞后，F10细胞分泌的PA增加，F1细胞和BL6细胞分泌的增加较少。此外，在PMA处理的F10细胞的条件培养基中检测到了第二种45,000道尔顿的纤溶酶原激活物活性。F10细胞和F1细胞的条件培养基中还显示含有一种33,000道尔顿的纤溶酶原激活物结合蛋白。经PMA处理后，F10细胞培养基中结合蛋白的浓度增加，而同样处理的F1细胞培养基中则没有增加。在对照和PMA处理的BL6细胞的条件培养基中几乎检测不到这种结合蛋白，很可能是一种纤溶酶原激活物抑制剂。因此，这三个在体内表型特征不同的亚系，在体外对蛋白酶和蛋白酶抑制剂合成的调节也不同。
