Membrane type-1-matrix metalloproteinase expressed by prostate carcinoma cells cleaves human laminin-5 beta3 chain and induces cell migration.
Membrane type-1-matrix metalloproteinase expressed by prostate carcinoma cells cleaves human laminin-5 beta3 chain and induces cell migration.
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发表时间：
2003-05
期刊：
影响因子：
11.2
通讯作者：
T. Udayakumar;Man-ling Chen;E. L. Bair;D. C. Von Bredow;A. Cress;R. Nagle;G. Bowden
中科院分区：
文献类型：
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作者：
T. Udayakumar;Man-ling Chen;E. L. Bair;D. C. Von Bredow;A. Cress;R. Nagle;G. Bowden
细胞外基质通过蛋白水解酶的降解是生理和病理组织重塑过程如滋养层细胞植入、伤口愈合和肿瘤侵袭的中心方面。我们假设前列腺腺癌细胞通过正常基底层的侵袭部分归因于金属蛋白酶诱导的层粘连蛋白-5（Ln-5）裂解和癌细胞运动性增强。我们研究了前列腺肿瘤细胞表面表达的膜1型基质金属蛋白酶（MT 1-MMP）在切割Ln-5和增强前列腺肿瘤细胞迁移中的作用。我们还确定了人类Ln-5的MT 1-MMP切割的性质以及这种改变的Ln-5如何改变前列腺癌细胞的迁移。我们发现人MT 1-MMP将纯化的人Ln-5切割成80-kDa片段。通过胰蛋白酶和胰凝乳蛋白酶对80-kDa裂解产物进行质谱分析，分别得到14个和9个不同的肽序列，其与Ln-5-β 3链的预期氨基酸序列相同。通过氨基酸计数，回收的肽代表Ln-5-β 3链的14.4%（胰蛋白酶）和10.3%（胰凝乳蛋白酶）。Ln-5的MT 1-MMP切割产物的胰蛋白酶和胰凝乳蛋白酶消化均未显示与Ln-5的其它链相同的任何其它肽。使用线性迁移试验，我们发现，由MT 1-MMP切割的Ln-5增强DU-145前列腺癌细胞的迁移2倍相比，未切割的Ln-5。使用封闭的反义MT 1-MMP寡核苷酸抑制DU-145细胞在Ln-5上的迁移。我们还发现，表达高水平MT 1-MMP的前列腺癌细胞，如PC 3 N和PPC，表现出增强的迁移在人Ln-5包被的基底上，并且这种迁移被阻断的反义MT 1-MMP寡核苷酸抑制。总之，这是一个新的和重要的发现，我们已经表明，β 3链被MT 1-MMP切割，这种切割增强前列腺癌细胞的迁移。
Degradation of the extracellular matrix by proteolytic enzymes is a central aspect of physiological and pathologic tissue-remodeling processes such as trophoblastic implantation, wound healing, and tumor invasion. We have hypothesized that prostate adenocarcinoma cell invasion through the normal basal lamina is attributable in part to metalloproteinase-induced cleavage of laminin-5 (Ln-5) and enhanced motility of the cancer cells. We studied the role of membrane type-1-matrix metalloproteinase (MT1-MMP) expressed on the surface of prostate tumor cells in cleaving Ln-5 and enhancing the migration of prostate tumor cells. We also determined the nature of the MT1-MMP cleavage of human Ln-5 and how this altered Ln-5 changes the migration of prostate carcinoma cells. We found that human MT1-MMP cleaves purified human Ln-5 to an 80-kDa fragment. Mass spectrometry analyses of the 80-kDa cleaved product by trypsin and chymotrypsin gave 14 and 9 different peptide sequences, respectively, that were identical to the expected amino acid sequence of the Ln-5-beta3 chain. The recovered peptides represent 14.4% (trypsin) and 10.3% (chymotrypsin) of Ln-5-beta3 chain by amino acid count. Both trypsin and chymotrypsin digestion of MT1-MMP-cleaved product of Ln-5 did not show any other peptides that were identical to the other chains of Ln-5. Using a linear migration assay we found that the Ln-5 cleaved by MT1-MMP enhanced the migration of DU-145 prostate carcinoma cells by 2-fold compared with uncleaved Ln-5. The use of blocked antisense MT1-MMP oligonucleotides inhibited the migration of DU-145 cells on Ln-5. We also found that the prostate carcinoma cells expressing high levels of MT1-MMP, such as PC3N and PPC, demonstrated enhanced migration on human Ln-5-coated substrate, and this migration was inhibited using blocked antisense MT1-MMP oligonucleotides. In conclusion, this is a novel and important finding where we have shown that beta3-chain is cleaved by MT1-MMP, and this cleavage enhances migration of prostate cancer cells.