Inhibition of cathepsin B and MMP-9 gene expression in glioblastoma cell line via RNA interference reduces tumor cell invasion, tumor growth and angiogenesis
Inhibition of cathepsin B and MMP-9 gene expression in glioblastoma cell line via RNA interference reduces tumor cell invasion, tumor growth and angiogenesis
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DOI：
10.1038/sj.onc.1207616
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发表时间：
2004-06-10
期刊：
影响因子：
8
通讯作者：
Rao, JS
中科院分区：
文献类型：
--
作者：
Lakka, SS;Gondi, CS;Rao, JS
细胞外蛋白水解酶通过蛋白水解级联反应协同影响基质的降解和肿瘤细胞的侵袭，单个蛋白水解酶在肿瘤的生长、侵袭、迁移和血管生成中起着不同的作用。基质金属蛋白酶-9和组织蛋白酶B参与了肿瘤的生长、血管形成和胶质瘤的侵袭过程。在本研究中，我们使用巨细胞病毒启动子驱动的DNA模板方法来诱导发夹RNA(HpRNA)触发的RNA干扰(RNAi)，从而通过单一的构建来阻断基质金属蛋白酶-9和组织蛋白酶B的基因表达。表达基质金属蛋白酶-9和组织蛋白酶B的双链RNA(DsRNA)表达载体显著抑制了基质金属蛋白酶-9和组织蛋白酶B的表达，并减少了SNB19胶质母细胞瘤细胞系在Matrigel和椭圆形侵袭模型中的侵袭行为。在体外和体内模型中，通过RNAi下调SNB19细胞中基质金属蛋白酶-9和组织蛋白酶B的表达，减少了人微血管内皮细胞之间的细胞间相互作用，导致毛细血管网络的形成中断。瘤内直接注射表达基质金属蛋白酶-9和组织蛋白酶B的hpRNA的DNA可显著抑制已建立的脑胶质瘤的生长和体内侵袭。进一步的腹腔注射(IP)表达hpRNA的基质金属蛋白酶-9和组织蛋白酶B的DNA在很长一段时间(4个月)内完全消退了预先建立的肿瘤，而没有任何这些肿瘤细胞的指征。这些观察首次表明，RNAi介导的同时靶向基质金属蛋白酶-9和组织蛋白酶B具有潜在的应用于人脑胶质瘤的治疗。
Extracellular proteases have been shown to cooperatively influence matrix degradation and tumor cell invasion through proteolytic cascades, with individual proteases having distinct roles in tumor growth, invasion, migration and angiogenesis. Matrix metalloproteases (MMP)-9 and cathepsin B have been shown to participate in the processes of tumor growth, vascularization and invasion of gliomas. In the present study, we used a cytomegalovirus promoter-driven DNA template approach to induce hairpin RNA (hpRNA)-triggered RNA interference (RNAi) to block MMP-9 and cathepsin B gene expression with a single construct. Transfection of a plasmid vector-expressing double-stranded RNA (dsRNA) for MMP-9 and cathepsin B significantly inhibited MMP-9 and cathepsin B expression and reduced the invasive behavior of SNB19, glioblastoma cell line in Matrigel and spheroid invasion models. Downregulation of MMP-9 and cathepsin B using RNAi in SNB19 cells reduced cell-cell interaction of human microvascular endothelial cells, resulting in the disruption of capillary network formation in both in vitro and in vivo models. Direct intratumoral injections of plasmid DNA expressing hpRNA for MMP-9 and cathepsin B significantly inhibited established glioma tumor growth and invasion in intracranial tumors in vivo. Further intraperitoneal (ip) injections of plasmid DNA expressing hpRNA for MMP-9 and cathepsin B completely regressed pre-established tumors for a long time (4 months) without any indication of these tumor cells. For the first time, these observations demonstrate that the simultaneous RNAi-mediated targeting of MMP-9 and cathepsin B has potential application for the treatment of human gliomas.