Interaction of human breast fibroblasts with collagen I increases secretion of procathepsin B
Interaction of human breast fibroblasts with collagen I increases secretion of procathepsin B
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DOI：
10.1074/jbc.m204708200
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发表时间：
2002-08-30
影响因子：
4.8
通讯作者：
Sloane, BF
中科院分区：
文献类型：
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作者：
Koblinski, JE;Dosescu, J;Sloane, BF
间质和肿瘤细胞与细胞外基质的相互作用可以调节蛋白酶的表达，包括溶酶体蛋白酶组织蛋白酶B和D。在本研究中，我们确定了这两种蛋白酶在人乳腺成纤维细胞中的表达是否受到与细胞外基质成分胶原I相互作用的调节。从非恶性乳腺组织以及恶性人乳腺肿瘤周围组织中分离乳腺成纤维细胞。这些成纤维细胞在I型胶原凝胶上的生长影响细胞形态，但不影响组织蛋白酶B或D染色的囊泡的细胞内定位。在胶原I凝胶或塑料上生长的成纤维细胞中，组织蛋白酶B和D水平（mRNA或细胞内蛋白）不受影响，这些细胞也不分泌组织蛋白酶D。相反，蛋白表达和分泌的组织蛋白酶B，主要是前组织蛋白酶B，诱导生长在胶原蛋白I凝胶。诱导的分泌似乎是由与胶原蛋白I结合的整联蛋白介导的，因为针对α（1）、α（2）和β（1）整联蛋白亚基的抑制性抗体阻止了胶原蛋白上生长的成纤维细胞分泌组织蛋白原B。此外，当细胞接种在β（1）整联蛋白抗体上时，可诱导组织蛋白酶原B分泌。据我们所知，这是第一次检查组织蛋白酶B和D在人乳腺成纤维细胞中的表达和定位及其由基质蛋白的调节。从乳腺成纤维细胞分泌半胱氨酸蛋白酶组织蛋白酶原B可能具有生理和病理后果，因为蛋白酶是乳腺正常发育和泌乳所需的，但也可以引发和加速乳腺癌的进展。
Interactions of stromal and tumor cells with the extracellular matrix may regulate expression of proteases including the lysosomal proteases cathepsins B and D. In the present study, we determined whether the expression of these two proteases in human breast fibroblasts was modulated by interactions with the extracellular matrix component, collagen I. Breast fibroblasts were isolated from non-malignant breast tissue as well as from tissue surrounding malignant human breast tumors. Growth of these fibroblasts on collagen I gels affected cell morphology, but not the intracellular localization of vesicles staining for cathepsin B or D. Cathepsins B and D levels (mRNA or intracellular protein) were not affected in fibroblasts growing on collagen I gels or plastic, nor was cathepsin D secreted from these cells. In contrast, protein expression and secretion of cathepsin B, primarily procathepsin B, was induced by growth on collagen I gels. The induced secretion appeared to be mediated by integrins binding to collagen I, as inhibitory antibodies against alpha(1), alpha(2), and beta(1) integrin subunits prevented procathepsin B secretion from fibroblasts grown on collagen. In addition, procathepsin B secretion was induced when cells were plated on beta(1) integrin antibodies. To our knowledge, this is the first examination of cathepsin B and D expression and localization in human breast fibroblasts and their regulation by a matrix protein. Secretion of the cysteine protease procathepsin B from breast fibroblasts may have physiological and pathological consequences, as proteases are required for normal development and for lactation of the mammary gland, yet can also initiate and accelerate the progression of breast cancer.