Tissue inhibitor of metalloproteinases-3 induces apoptosis in melanoma cells by stabilization of death receptors
Tissue inhibitor of metalloproteinases-3 induces apoptosis in melanoma cells by stabilization of death receptors
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DOI：
10.1038/sj.onc.1206292
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发表时间：
2003-04-10
期刊：
影响因子：
8
通讯作者：
Kähäri, VM
中科院分区：
文献类型：
--
作者：
Ahonen, M;Poukkula, M;Kähäri, VM
金属蛋白酶组织抑制剂（TIMPs）是基质金属蛋白酶（MMP）和adamalysin （ADAM）活性的重要调节因子。我们之前已经证明腺病毒表达的组织金属蛋白酶抑制剂-3 （TIMP-3）诱导黑色素瘤细胞凋亡并抑制人类黑色素瘤异种移植物的生长。在此，我们研究了死亡受体在TIMP-3诱导的黑色素瘤细胞凋亡中的作用。我们的研究结果表明，三种转移性黑色素瘤细胞系（A2058、SK-Mel-5和WM-266-4）暴露于重组TIMP-3、TIMP-3 n端MMP抑制结构域以及腺病毒表达的TIMP-3，可稳定黑色素瘤细胞表面的肿瘤坏死因子受体-1 （TNF-RI）、FAS和tnf相关凋亡诱导配体受体-1 (TRAIL- ri)，并使这些细胞对tnf - α、抗FAS抗体和TRAIL诱导的凋亡敏感。TIMP-3稳定死亡受体导致caspase-8和caspase-3的激活，随后的凋亡被特异性caspase-8抑制剂（Z-IETD-FMK）和泛caspase抑制剂（Z-DEVD-FMK）阻断。在体内，腺病毒介导的TIMP-3在人黑色素瘤异种移植物中的表达导致TNF-RI、FAS和cleaved caspase-3的免疫染色增加，并导致黑色素瘤细胞凋亡。综上所述，这些结果表明TIMP-3通过稳定三种不同的死亡受体和通过caspase-8激活它们的凋亡信号级联来促进黑色素瘤细胞的凋亡。
Tissue inhibitors of metalloproteinases (TIMPs) are important regulators of matrix metalloproteinase (MMP) and adamalysin (ADAM) activity. We have previously shown that adenovirally expressed tissue inhibitor of metalloproteinases-3 (TIMP-3) induces apoptosis in melanoma cells and inhibits growth of human melanoma xenografts. Here, we have studied the role of death receptors in apoptosis of melanoma cells induced by TIMP-3. Our results show, that the exposure of three metastatic melanoma cell lines (A2058, SK-Mel-5, and WM-266-4) to recombinant TIMP-3, N-terminal MMP inhibitory domain of TIMP-3, as well as to adenovirally expressed TIMP-3 results in stabilization of tumor necrosis factor receptor-1 (TNF-RI), FAS, and TNF-related apoptosis inducing ligand receptor-1 (TRAIL-RI) on melanoma cell surface and sensitizes these cells to apoptosis induced by TNF-alpha, anti-Fas-antibody and TRAIL. Stabilization of death receptors by TIMP-3 results in activation of caspase-8 and caspase-3, and subsequent apoptosis is blocked by specific caspase-8 inhibitor (Z-IETD-FMK) and by pan-caspase inhibitor (Z-DEVD-FMK). Adenovirus-mediated expression of TIMP-3 in human melanoma xenografts in vivo resulted in increased immunostaining for TNF-RI, FAS, and cleaved caspase-3, and in apoptosis of melanoma cells. Taken together, these results show that TIMP-3 promotes apoptosis in melanoma cells through stabilization of three distinct death receptors and activation of their apoptotic signaling cascade through caspase-8.