microRNA-9 Targets Matrix Metalloproteinase 14 to Inhibit Invasion, Metastasis, and Angiogenesis of Neuroblastoma Cells
microRNA-9 Targets Matrix Metalloproteinase 14 to Inhibit Invasion, Metastasis, and Angiogenesis of Neuroblastoma Cells
复制标题
microRNA-9 靶向基质金属蛋白酶 14 抑制神经母细胞瘤细胞的侵袭、转移和血管生成
DOI：
10.1158/1535-7163.mct-12-0001
复制
发表时间：
2012-07-01
影响因子：
5.7
通讯作者：
Tong, Qiangsong
中科院分区：
文献类型：
--
作者：
Zhang, Huanyu;Qi, Meng;Tong, Qiangsong
基质金属蛋白酶(MMP14)是唯一一种在肿瘤转移和血管生成中起关键作用的膜锚定的MMPs。然而，在肿瘤中表达基质金属蛋白酶-14的机制仍然很大程度上仍不清楚。在本研究中，有29/42例神经母细胞瘤组织中检测到了基质金属蛋白酶-14的表达，其表达与神经母细胞瘤的临床病理特征和患者的生存期有关。在20例神经母细胞瘤中，microRNA9(miR-9)表达下调，且与MMP14的表达呈负相关。生物信息学分析显示，在MMP14mRNA的3‘-非翻译区(3’-UTR)有一个miR-9结合位点。在培养的神经母细胞瘤细胞系SH-SY5Y和SK-N-SH中，miR-9的过表达或基因敲除反应地改变了基质金属蛋白酶-14及其下游基因血管内皮生长因子的mRNA和蛋白水平。在基质金属蛋白酶-14 3‘-非编码区荧光素酶报告系统中，miR-9下调荧光素酶活性，这种作用被miR-9结合位点的突变所消除。在体内外，miR-9过表达抑制SH-SY5Y和SK-N-SH细胞的侵袭、转移和血管生成。此外，miR-9对基质金属蛋白酶-14的表达、黏附、迁移、侵袭和血管生成的影响通过在这些细胞中的过表达而得以挽救。此外，抗miR-9抑制剂或敲除MMP14分别增加或抑制神经母细胞瘤细胞的迁移、侵袭和血管生成。这些结果表明miR-9通过3‘-非编码区的结合部位抑制了基质金属蛋白酶-14的表达，从而抑制了神经母细胞瘤的侵袭、转移和血管生成。摩尔癌症治疗；11(7)；1454-66。©2012 AACR。
Matrix metalloproteinase (MMP)-14 is the only membrane-anchored MMP that plays a critical role in tumor metastasis and angiogenesis. However, the mechanisms underlying MMP-14 expression in tumors still remain largely unknown. In this study, MMP-14 immunostaining was identified in 29/42 neuroblastoma tissues, which was correlated with clinicopathologic features and shorter patients' survival. In subtotal 20 neuroblastoma cases, microRNA 9 (miR-9) was downregulated and inversely correlated with MMP-14 expression. Bioinformatics analysis revealed a putative miR-9–binding site in the 3′-untranslated region (3′-UTR) of MMP-14 mRNA. Overexpression or knockdown of miR-9 responsively altered both the mRNA and protein levels of MMP-14 and its downstream gene, vascular endothelial growth factor, in cultured neuroblastoma cell lines SH-SY5Y and SK-N-SH. In an MMP-14 3′-UTR luciferase reporter system, miR-9 downregulated the luciferase activity, and these effects were abolished by a mutation in the putative miR-9–binding site. Overexpression of miR-9 suppressed the invasion, metastasis, and angiogenesis of SH-SY5Y and SK-N-SH cells in vitro and in vivo. In addition, the effects of miR-9 on MMP-14 expression, adhesion, migration, invasion, and angiogenesis were rescued by overexpression of MMP-14 in these cells. Furthermore, anti-miR-9 inhibitor or knockdown of MMP-14 respectively increased or inhibited the migration, invasion, and angiogenesis of neuroblastoma cells. These data indicate that miR-9 suppresses MMP-14 expression via the binding site in the 3′-UTR, thus inhibiting the invasion, metastasis, and angiogenesis of neuroblastoma. Mol Cancer Ther; 11(7); 1454–66. ©2012 AACR.