Departamento de Bioquímica, Universidade Federal de São Paulo-Escola Paulista de Medicina, Rua Três de Maio 100, 04044-020, São Paulo, SP, Brazil.
Biol Chem. 2011 Apr;392(4):327-36. doi: 10.1515/BC.2011.031.
Supplementary to the efficient inhibition of trypsin, chymotrypsin, plasma kallikrein, and plasmin already described by the EcTI inhibitor from Enterolobium contortisiliquum, it also blocks human neutrophil elastase (K(iapp)=4.3 nM) and prevents phorbol ester (PMA)-stimulated activation of matrix metalloproteinase (MMP)-2 probably via interference with membrane-type 1 (MT1)-MMP. Moreover, plasminogen-induced activation of proMMP-9 and processing of active MMP-2 was also inhibited. Furthermore, the effect of EcTI on the human cancer cell lines HCT116 and HT29 (colorectal), SkBr-3 and MCF-7 (breast), K562 and THP-1 (leukemia), as well as on human primary fibroblasts and human mesenchymal stem cells (hMSCs) was studied. EcTI inhibited in a concentration range of 1.0-2.5 μM rather specifically tumor cell viability without targeting primary fibroblasts and hMSCs. Taken together, our data indicate that the polyspecific proteinase inhibitor EcTI prevents proMMP activation and is cytotoxic against tumor cells without affecting normal tissue remodeling fibroblasts or regenerative hMSCs being an important tool in the studies of tumor cell development and dissemination.
除了已被描述的高效抑制胰蛋白酶、糜蛋白酶、血浆激肽释放酶和纤溶酶之外，来自皱荚黎豆的 EcTI 抑制剂还能抑制人中性粒细胞弹性蛋白酶（K(i app )=4.3 nM），并通过干扰膜型 1（MT1）-MMP 来防止佛波酯（PMA）刺激的基质金属蛋白酶（MMP）-2 的激活。此外，纤溶酶原诱导的 proMMP-9 的激活和活性 MMP-2 的加工也被抑制。此外，还研究了 EcTI 对人癌细胞系 HCT116 和 HT29（结直肠）、SkBr-3 和 MCF-7（乳腺）、K562 和 THP-1（白血病）以及人原代成纤维细胞和人间充质干细胞（hMSC）的影响。EcTI 在 1.0-2.5 μM 的浓度范围内抑制肿瘤细胞活力，而对原代成纤维细胞和 hMSC 的靶向作用较小。总之，我们的数据表明，多特异性蛋白酶抑制剂 EcTI 可防止 proMMP 的激活，并对肿瘤细胞具有细胞毒性，而不会影响正常组织重塑的成纤维细胞或再生的 hMSC，是研究肿瘤细胞发展和扩散的重要工具。
