Degradation of basement membrane type IV collagen and lung subendothelial matrix by rat mammary adenocarcinoma cell clones of differing metastatic potentials.
Degradation of basement membrane type IV collagen and lung subendothelial matrix by rat mammary adenocarcinoma cell clones of differing metastatic potentials.
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发表时间：
1987-09
期刊：
影响因子：
11.2
通讯作者：
M. Nakajima;D. Welch;P. Belloni;G. Nicolson
中科院分区：
文献类型：
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作者：
M. Nakajima;D. Welch;P. Belloni;G. Nicolson
一系列的大鼠13762 NF乳腺癌细胞克隆和亚克隆的各种肺转移的潜力进行了检查，其能力，以降解大鼠肺内皮下基质和纯化的基底膜IV型胶原。同时根据s.c.注射或静脉内注射细胞后的“实验性”肺转移。体外培养大鼠肺微血管内皮细胞，用[3 H]脯氨酸代谢标记含有IV型胶原的内皮下基质。当乳腺癌细胞被放置在隔离的内皮下基质，碎片和溶解的[3 H]脯氨酸标记的组件进行观察，高转移性13762 NF细胞溶解的矩阵在更高的速率比转移性差的细胞。使用从Engelbreth-Holm-Swarm肿瘤中纯化的[3 H]脯氨酸标记的IV型胶原作为底物，测定13762 NF细胞的IV型胶原溶解活性。我们发现IV型胶原溶解活性的活细胞和他们的“自发性”肺转移潜能之间有很好的相关性（r = 0.993）。条件培养基中IV型胶原溶解活性的水平取决于细胞培养条件。在酸处理的胎牛血清的存在或不存在下，高转移性细胞分泌的IV型胶原溶解酶的活性和潜伏形式比转移性差的细胞更高的量。将IV型前胶原与由高转移性13762 NF细胞调节并用胰蛋白酶处理的培养基一起孵育，导致产生几个IV型胶原特征性的大片段。结果表明，基底膜IV型胶原的酶降解是重要的13762 NF乳腺癌细胞的肺转移。
A series of rat 13762NF mammary adenocarcinoma cell clones and subclones of various lung metastatic potentials were examined for their abilities to degrade rat lung subendothelial matrix and purified basement membrane type IV collagen. Metastatic potentials were simultaneously determined based on the ability to form "spontaneous" lung metastases after s.c. injection or "experimental" lung metastases after i.v. injection of cells. Microvessel endothelial cells isolated from rat lung were grown in vitro, and the subendothelial matrix containing type IV collagen was metabolically labeled with [3H]proline. When mammary adenocarcinoma cells were placed on the isolated subendothelial matrix, fragmentation and solubilization of [3H]proline-labeled components were observed; highly metastatic 13762NF cells solubilized the matrix at higher rates than did poorly metastatic cells. The 13762NF cells were assayed for type IV collagenolytic activity using [3H]proline-labeled type IV collagen purified from Engelbreth-Holm-Swarm tumor as a substrate. We found excellent correlation between the type IV collagenolytic activities of living cells and their "spontaneous" lung metastatic potentials (r = 0.993). The levels of type IV collagenolytic activity in the conditioned medium depended on the cell culture conditions. In the presence or absence of acid-treated fetal bovine serum, highly metastatic cells secreted higher amounts of type IV collagenolytic enzymes in active and latent forms than did poorly metastatic cells. Incubation of procollagen type IV with medium conditioned by highly metastatic 13762NF cells and treated with trypsin resulted in the production of several large fragments characteristic of type IV collagen. The results suggest that enzymatic degradation of basement membrane type IV collagen is important in lung metastasis of 13762NF mammary adenocarcinoma cells.