SECRETION OF EXTRACELLULAR MATRIX-DEGRADING PROTEINASES IS INCREASED IN EPITHELIAL OVARIAN-CARCINOMA
SECRETION OF EXTRACELLULAR MATRIX-DEGRADING PROTEINASES IS INCREASED IN EPITHELIAL OVARIAN-CARCINOMA
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DOI：
10.1002/ijc.2910560415
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发表时间：
1994-02-15
影响因子：
6.4
通讯作者：
STACK, MS
中科院分区：
文献类型：
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作者：
MOSER, TL;YOUNG, TN;STACK, MS
与肿瘤侵袭相关的生化事件包括肿瘤相关蛋白对基底膜的局部降解。在本研究中，我们研究了5个卵巢上皮性癌细胞株(DOV 13、OVCA 420、OVCA 429、OVCA 432、OVCA 433)以及正常卵巢上皮细胞的蛋白水解酶分泌谱。免疫捕获试验表明，5株癌细胞均可产生分泌型和表面相关的纤溶酶原激活剂。尿型纤溶酶原激活物(u-PA)的产量比组织型纤溶酶原激活物(t-PA)的产量高一个数量级。此外，正常卵巢上皮细胞不能检测到t-PA的分泌，而u-PA的产生比卵巢癌细胞低17-38倍。Western-blotting分析表明，在无血清培养条件下，u-PA以单链形式(suc-PA)形式分泌。将纤溶酶原与卵巢癌细胞条件培养液孵育，可使酶原直接激活为纤溶酶。此外，在细胞与纤溶酶原孵育后，纤溶酶从细胞表面洗脱出来，表明卵巢癌细胞含有纤溶酶原/纤溶酶的结合部位，表面相关的纤溶酶原激活剂可以接触到这些结合部位。除纤溶酶原激活剂外，DOV-13、OVCA-429和OVCA-433细胞也产生金属蛋白酶。DOV 13细胞产生一种类似于基质金属蛋白酶2(MMP2)的68 kDa的金属蛋白酶，而OVCA429和433分泌一种类似于MMP9的92 kDa的酶。卵巢癌相关纤溶酶原激活物和金属蛋白酶共同催化主要基底膜蛋白组分、IV型胶原、IV型明胶、层粘连蛋白和纤维连接蛋白的水解。卵巢癌细胞相对于正常卵巢上皮具有较强的蛋白分解能力，提示卵巢上皮癌的侵袭和扩散可能是通过一种生化机制来调节的。(C)1994年Wiley-Liss，Inc.
The biochemical events associated with tumor invasion involve localized degradation of the basement membrane by tumor-associated proteinases. In this study, we have characterized the proteinase secretion profiles of 5 ovarian epithelial carcinoma cell lines (DOV 13, OVCA 420, OVCA 429, OVCA 432, OVCA 433) as well as normal ovarian epithelial cells. Immunocapture assays demonstrated that all 5 carcinoma cell lines produce both secreted and surface-associated plasminogen activator. Urinary-type plasminogen activator (u-PA) production was one order of magnitude greater than production of tissue-type plasminogen activator (t-PA). Furthermore, t-PA secretion by normal ovarian epithelial cells was not detectable, whereas u-PA production was 17- to 38-fold lower than in ovarian carcinoma cells. Western-blotting analysis demonstrated that u-PA was secreted as the single chain form (suc-PA) when cells were cultured in serum-free medium. Incubation of plasminogen with ovarian carcinoma cell-conditioned medium resulted in direct activation of the zymogen to plasmin. Furthermore, following incubation of cells with plasminogen, plasmin was eluted from the cell surface, indicating that ovarian carcinoma cells contain binding sites for plasminogen/plasmin which are accessible to surface-associated plasminogen activators. In addition to plasminogen activators, metallo-proteinases were also produced by DOV 13, OVCA 429 and OVCA 433 cells. DOV 13 cells produce a 68-kDa metalloproteinase similar to matrix metalloproteinase 2 (MMP-2) whereas a 92-kDa enzyme similar to MMP-9 is secreted by OVCA 429 and 433. Together, ovarian carcinoma-associated plasminogen activators and metalloproteinases catalyze the hydrolysis of the major basement membrane protein components, type-iv collagen, type-iv gelatin, laminin and fibronectin. The enhanced proteolytic capability of ovarian carcinoma cells relative to normal ovarian epithelium suggests a biochemical mechanism by which invasion and spread of ovarian epithelial carcinoma may be mediated. (C) 1994 Wiley-Liss, Inc.