Cathepsin D Specifically Cleaves the Chemokines Macrophage Inflammatory Protein-1α, Macrophage Inflammatory Protein-1β, and SLC That Are Expressed in Human Breast Cancer
Cathepsin D Specifically Cleaves the Chemokines Macrophage Inflammatory Protein-1α, Macrophage Inflammatory Protein-1β, and SLC That Are Expressed in Human Breast Cancer
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DOI：
10.1016/s0002-9440(10)63914-4
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发表时间：
2003-04
影响因子：
6
通讯作者：
M. Wolf;I. Clark‐Lewis;C. Buri;H. Langen;M. Lis;L. Mazzucchelli
中科院分区：
文献类型：
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作者：
M. Wolf;I. Clark‐Lewis;C. Buri;H. Langen;M. Lis;L. Mazzucchelli
组织蛋白酶D （Cath-D）在人类原发性乳腺癌中的表达与预后不良有关。为了更好地了解可能参与癌症侵袭和转移的Cath-D底物，我们研究了这种蛋白酶和趋化因子之间的潜在相互作用。在这里，我们报道纯化的Cath-D，以及从人乳腺癌细胞系MCF-7和T47D培养的上清，选择性地降解巨噬细胞炎症蛋白(MIP)-1α (CCL3), MIP-1β (CCL4)和SLC （CCL21）。蛋白酶抑制剂pepstatin A完全阻断了蛋白水解，并且使用大型趋化因子面板证明了Cath-D切割的特异性。MIP-1α和MIP-1β的降解是快速和完全的，而SLC的裂解是缓慢和不完全的。质谱分析表明，Cath-D可切割SLC的leu58 - trp59键，产生两个功能无活性片段。对一系列单核细胞趋化蛋白-3/MIP-1β杂交体的cat - d蛋白水解分析表明，MIP-1β的加工可能从切割位于c端结构域的氨基酸开始。原位杂交研究发现MIP-1α、MIP-1β和Cath-D基因主要在乳腺癌的间质室中表达，而SLC转录本则在肿瘤组织的毛细血管和小静脉内皮细胞中发现。通过对培养上清和细胞裂解物进行酶联免疫吸附试验评估，乳腺癌细胞系MCF-7和T47D中Cath-D的产生不受白细胞介素-8 （CXCL8）、SDF-1 （CXCL12）和SLC等趋化因子刺激的影响。这些数据表明，通过cat - d使趋化因子失活可能影响肿瘤细胞外微环境中的调节机制，进而可能影响抗肿瘤免疫应答的产生、癌细胞的迁移，或两者兼而有之。
Cathepsin D (Cath-D) expression in human primary breast cancer has been associated with a poor prognosis. In search of a better understanding of the Cath-D substrates possibly involved in cancer invasiveness and metastasis, we investigated the potential interactions between this protease and chemokines. Here we report that purified Cath-D, as well as culture supernatants from the human breast carcinoma cell lines MCF-7 and T47D, selectively degrade macrophage inflammatory protein (MIP)-1α (CCL3), MIP-1β (CCL4), and SLC (CCL21). Proteolysis was totally blocked by the protease inhibitor pepstatin A, and specificity of Cath-D cleavage was demonstrated using a large chemokine panel. Whereas MIP-1α and MIP-1β degradation was rapid and complete, cleavage of SLC was slow and not complete. Mass spectrometry analysis showed that Cath-D cleaves the Leu58to Trp59bond of SLC producing two functionally inactive fragments. Analysis of Cath-D proteolysis of a series of monocyte chemoattractant protein-3/MIP-1β hybrids indicated that processing of MIP-1β might start by cleaving off amino acids located in the C-terminal domain. In situ hybridization studies revealed MIP-1α, MIP-1β, and Cath-D gene expression mainly in the stromal compartment of breast cancers whereas SLC transcripts were found in endothelial cells of capillaries and venules within the neoplastic tissues. Cath-D production in the breast carcinoma cell lines MCF-7 and T47D, as assessed by enzyme-linked immunosorbent assay of culture supernatants and cell lysates, was not affected by stimulation with chemokines such as interleukin-8 (CXCL8), SDF-1 (CXCL12), and SLC. These data suggest that inactivation of chemokines by Cath-D possibly influences regulatory mechanisms in the tumoral extracellular microenvironment that in turn may affect the generation of the antitumoral immune response, the migration of cancer cells, or both processes.