Membrane type I-matrix metalloproteinase-mediated degradation of type I collagen by oral squamous cell carcinoma cells.
Membrane type I-matrix metalloproteinase-mediated degradation of type I collagen by oral squamous cell carcinoma cells.
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发表时间：
2001-08
期刊：
影响因子：
11.2
通讯作者：
S. Aznavoorian;Bryan A. Moore;L. D. Alexander-Lister;S. Hallit;L. Windsor;Jeffrey A. Engler
中科院分区：
文献类型：
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作者：
S. Aznavoorian;Bryan A. Moore;L. D. Alexander-Lister;S. Hallit;L. Windsor;Jeffrey A. Engler
口腔鳞状细胞癌是一种高度侵袭性的病变，破坏邻近组织并侵入骨骼和肌肉，这很可能是基质金属蛋白酶（MMP）活性的结果。我们研究了三种来源于舌鳞状细胞癌的细胞系降解间质胶原的内在能力，目的是确定基质降解酶。SCC-25和SCC-15细胞在不存在外源性生长因子或细胞因子的情况下，当作为集落接种在干膜上时，降解重构的纤维状I型胶原。降解仅限于皮下基质，被佛波酯增强2-3倍，并且是严格MMP依赖性的，因为它被BB-94和金属蛋白酶-2的组织抑制剂阻断，但不被丝氨酸和半胱氨酸蛋白酶的抑制剂阻断。通过表面生物素化和免疫沉淀检测，两种细胞系在其表面上表达活性（M（r）57，000）膜型I-MMP（MT 1-MMP）。同时，这两种细胞系激活内源性MMP-2时，培养在I型胶原膜，如通过酶谱法评估。佛波酯处理增强胶原诱导的MMP-2活化，这伴随着表面标记的M（r）43，000形式的MT 1-MMP的出现。用合成弗林蛋白酶抑制剂处理细胞，其抑制MT 1-MMP酶原的加工，阻断胶原降解。相反，CAL 27细胞在基础或佛波醇12-肉豆蔻酸酯13-乙酸酯刺激条件下不降解胶原。尽管通过免疫印迹分析在这些细胞中可检测到proMT 1-MMP（M（r）63，000/65，000），但相对于SCC-25和SCC-15细胞，它们在其表面上表达的活性MT 1-MMP水平大大降低。相应地，在胶原上培养的CAL 27细胞既不表达潜伏的也不表达活性明胶酶。裂解物和条件培养基的免疫印迹揭示了proMMP-1和proMMP-13在所有三种细胞系中的组成型表达。我们的结论是，在没有外源性生长因子或辅助基质细胞，口腔鳞状细胞癌细胞降解间质胶原蛋白需要一个阈值水平的活性MT 1-MMP细胞表面。
Oral squamous cell carcinomas are highly invasive lesions that destroy adjacent tissues and invade bone and muscle, which is most likely the result of matrix metalloproteinase (MMP) activity. We examined three cell lines derived from squamous cell carcinoma of the tongue for their intrinsic capacities to degrade interstitial collagen with the goal of identifying the matrix-degrading enzymes. SCC-25 and SCC-15 cells degrade reconstituted fibrillar type I collagen in the absence of exogenous growth factors or cytokines when seeded as a colony on dried films. Degradation is confined to the subjacent matrix, is enhanced 2-3-fold by phorbol ester, and is strictly MMP-dependent, as it is blocked by BB-94 and tissue inhibitor of metalloproteinases-2 but not by inhibitors of serine and cysteine proteinases. Both cell lines express active (M(r) 57,000) membrane type I-MMP (MT1-MMP) on their surfaces, as detected by surface biotinylation and immunoprecipitation. Concomitantly, both cell lines activate endogenous MMP-2 when cultured on type I collagen films, as assessed by zymography. Phorbol ester treatment enhances collagen-induced MMP-2 activation, which is accompanied by the appearance of a surface-labeled M(r) 43,000 form of MT1-MMP. Treatment of cells with a synthetic furin inhibitor, which inhibits processing of the MT1-MMP zymogen, blocks collagen degradation. In contrast, CAL 27 cells do not degrade collagen under either basal or phorbol 12-myristate 13-acetate-stimulated conditions. Although proMT1-MMP (M(r) 63,000/65,000) is detectable in these cells by immunoblot analysis, they express greatly reduced levels of active MT1-MMP on their surfaces relative to SCC-25 and SCC-15 cells. Correspondingly, CAL 27 cells cultured on collagen express neither latent nor active gelatinases. Immunoblots of lysates and conditioned media revealed the constitutive expression of proMMP-1 and proMMP-13 in all three cell lines. We conclude that in the absence of exogenous growth factors or accessory stromal cells, degradation of interstitial collagen by oral squamous cell carcinoma cells requires a threshold level of active MT1-MMP on cell surfaces.