INHIBITORY EFFECT OF A CONJUGATE BETWEEN HUMAN UROKINASE AND URINARY TRYPSIN-INHIBITOR ON TUMOR-CELL INVASION IN-VITRO
INHIBITORY EFFECT OF A CONJUGATE BETWEEN HUMAN UROKINASE AND URINARY TRYPSIN-INHIBITOR ON TUMOR-CELL INVASION IN-VITRO
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DOI：
10.1074/jbc.270.14.8361
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发表时间：
1995-04-07
影响因子：
4.8
通讯作者：
TERAO, T
中科院分区：
文献类型：
--
作者：
KOBAYASHI, H;GOTOH, J;TERAO, T
蛋白水解酶如尿激酶型纤溶酶原激活剂（uPA）、纤溶酶和胶原酶介导多种肿瘤细胞的蛋白水解。肿瘤细胞分泌的uPA可以通过uPA分子的氨基末端片段（ATF）内的生长因子样结构域与细胞表面受体结合，具有高亲和力。尿胰蛋白酶抑制剂（UTI）能有效抑制可溶性和肿瘤细胞表面受体结合的纤溶酶，从而减少肿瘤细胞的侵袭和转移的形成。抗侵袭作用取决于UTI分子的抗纤溶酶活性，特别是结构域II。我们合成了人uPA的ATF与UTI分子或UTI结构域II （HI-8）的偶联物。共轭物（ATF）的作用。UTI或ATF。研究了HI-8对肿瘤细胞体外侵袭的影响。ATF。UTI和ATF。HI-8以快速、饱和、剂量依赖性和可逆的方式结合到U937细胞上。受体结合ATF的很大一部分。UTI和ATF。37℃下，HI-8在细胞表面停留至少5小时。ATF抑制肿瘤细胞表面受体结合的纤溶酶。UTI和ATF。与ATF、UTI或HI-8处理的肿瘤细胞相比，HI-8明显增强。细胞侵袭试验结果显示，ATF。UTI和ATF。HI-8对表达uPA受体的肿瘤细胞特异性靶向HI-8非常有效，而缺乏uPA受体的肿瘤细胞可能受其偶联物的影响较小。我们的研究结果表明，细胞表面uPA和纤溶酶活性对侵袭过程至关重要，并且结合物对细胞表面的紧密环境表现出纤溶酶抑制作用，随后通过Matrigel在体外侵袭实验中抑制肿瘤细胞的侵袭。
Proteolytic enzymes such as urokinase-type plasminogen activator (uPA), plasmin, and collagenase mediate proteolysis by a variety of tumor cells. uPA secreted by tumor cells can be bound to a cell surface receptor via a growth factor-like domain within the amino-terminal fragment (ATF) of the uPA molecule with high affinity. Urinary trypsin inhibitor (UTI) efficiently inhibits the soluble and the tumor cell-surface receptor-bound plasmin and subsequently reduces tumor cell invasion and the formation of metastasis. The anti-invasive effect is dependent on the anti-plasmin activity of the UTI molecule, domain II in particular. We synthesized a conjugate between ATF of human uPA and a native UTI molecule or domain II of UTI (HI-8). The effect of the conjugates (ATF . UTI or ATF . HI-8) on tumor cell invasion in vitro was investigated. ATF . UTI and ATF . HI-8 bound to U937 cells in a rapid, saturable, dose-dependent, and reversible manner. A large part of receptor-bound ATF . UTI and ATF . HI-8 remains on the cell surface for at least 5 h at 37 degrees C. Inhibition of tumor cell surface receptor-bound plasmin by ATF . UTI and ATF . HI-8 was markedly enhanced when compared with tumor cells treated either with ATF, UTI, or HI-8. Results of a cell invasion assay showed that ATF . UTI and ATF . HI-8 is very effective at targeting HI-8 specifically to uPA receptor-expressing tumor cells, whereas tumor cells devoid of uPA receptor may be less affected by the conjugates. Our results indicate that cell surface uPA and plasmin activity is essential to the invasive process and that the conjugates exhibit plasmin inhibition to the close environment of the cell surface and subsequently inhibit the tumor cell invasion through Matrigel in an in vitro invasion assay.