Klisovic Dino D, Klisovic Marko I, Effron David, Liu Shujun, Marcucci Guido, Katz Steven E
William H. Havener Eye Center, Columbus, OH, USA.
Melanoma Res. 2005 Jun;15(3):147-53. doi: 10.1097/00008390-200506000-00002.
Uveal melanoma (UM) is a highly malignant primary intraocular tumour in adults that has a high mortality rate due to haematogenous dissemination. The migration of UM cells through the basement membrane requires the presence of proteolytic enzymes, such as matrix metalloproteinases (MMPs). The expression of MMP-2, MMP-9 and membrane type-1/MMP (MT-1/MMP) in UM cells is a known risk factor for metastatic disease. We tested the effect of depsipeptide (DP) on UM cell migration and the level and activity of MMP-2, MMP-9, MT-1/MMP and tissue inhibitors of matrix metalloproteinases 1 and 2 (TIMP-1 and TIMP-2). Three primary and two metastatic (liver metastasis) UM cell lines were treated with DP (0, 1, 5 and 10 nmol/l) for 24 h. Migration of UM cells was studied in modified Boyden migration chambers for 24 h and only viable cells on both sides of the membrane were counted. Enzyme-linked immunosorbent assays (ELISAs) were used to quantify the level of MMP-2, MMP-9, MT-1/MMP, TIMP-1 and TIMP-2 after the cells had been exposed to DP (0, 1, 5 and 10 nmol/l) for 24 h. In addition, the activities of MMP-2, MMP-9 and MT-1/MMP were determined after DP treatment. A dose-dependent decrease in the migration of viable UM cells was observed for primary and metastatic cell lines (30-50% inhibition). We detected a dose-dependent: (1) decrease in the protein level of MMP-2, MMP-9 and MT-1/MMP; (2) decrease in the activity of MMP-2, MMP-9 and MT-1/MMP; and (3) increase in the protein level of TIMP-1 and TIMP-2. It can be concluded that DP is a potent inhibitor of primary and metastatic UM cell migration in vitro. Our data suggest that this inhibition is mediated by the downregulation of MMPs and the upregulation of TIMPs. DP may be a valuable adjunctive treatment modality for primary and metastatic UM in humans.
葡萄膜黑色素瘤（UM）是成人中一种高度恶性的原发性眼内肿瘤，由于血行播散，其死亡率很高。UM细胞通过基底膜的迁移需要蛋白水解酶的存在，如基质金属蛋白酶（MMPs）。UM细胞中MMP-2、MMP-9和膜型-1/MMP（MT-1/MMP）的表达是转移性疾病的已知危险因素。我们测试了缩肽（DP）对UM细胞迁移以及MMP-2、MMP-9、MT-1/MMP和基质金属蛋白酶组织抑制剂1和2（TIMP-1和TIMP-2）水平及活性的影响。用DP（0、1、5和10 nmol/l）处理三种原发性和两种转移性（肝转移）UM细胞系24小时。在改良的博伊登迁移小室中研究UM细胞的迁移24小时，仅计数膜两侧的活细胞。在细胞暴露于DP（0、1、5和10 nmol/l）24小时后，使用酶联免疫吸附测定（ELISA）来定量MMP-2、MMP-9、MT-1/MMP、TIMP-1和TIMP-2的水平。此外，在DP处理后测定MMP-2、MMP-9和MT-1/MMP的活性。对于原发性和转移性细胞系，观察到活UM细胞迁移呈剂量依赖性降低（抑制率为30 - 50%）。我们检测到剂量依赖性：（1）MMP-2、MMP-9和MT-1/MMP蛋白水平降低；（2）MMP-2、MMP-9和MT-1/MMP活性降低；（3）TIMP-1和TIMP-2蛋白水平升高。可以得出结论，DP是体外原发性和转移性UM细胞迁移的有效抑制剂。我们的数据表明，这种抑制是由MMPs的下调和TIMPs的上调介导的。DP可能是人类原发性和转移性UM的一种有价值的辅助治疗方式。
