RNA interference-mediated simultaneous down-regulation of urokinase-type plasminogen activator receptor and cathepsin B induces caspase-8-mediated apoptosis in SNB19 human glioma cells
RNA interference-mediated simultaneous down-regulation of urokinase-type plasminogen activator receptor and cathepsin B induces caspase-8-mediated apoptosis in SNB19 human glioma cells
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DOI：
10.1158/1535-7163.mct-05-0531
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发表时间：
2006-12-01
影响因子：
5.7
通讯作者：
Rao, Jasti S.
中科院分区：
文献类型：
--
作者：
Gondi, Christopher S.;Kandhukuri, Neelima;Rao, Jasti S.
胶质瘤的侵袭性取决于周围细胞外基质的蛋白水解裂解。组织蛋白酶B和尿激酶型纤溶酶原激活物受体（uPAR）一起在胶质瘤中过度表达，因此是基因治疗的有吸引力的靶点。在本研究中，我们使用质粒构建诱导RNA干扰（RNAi）介导的uPAR和组织蛋白酶B在SNB19人胶质瘤细胞中的下调。我们观察到uPAR和组织蛋白酶B的同时下调诱导了促凋亡基因的上调，并引发了线粒体Delta psi的崩溃。组织蛋白酶B和uPAR下调的细胞活化caspase-8和DFF40/caspase-活化DNase的表达增加。还观察到AIF的核易位和Fas配体向细胞膜的易位。Ki67和X-linked inhibitor蛋白水平下降，提示凋亡。这些结果提示细胞表面的upar -组织蛋白酶B复合物参与维持SNB19胶质瘤细胞的活力。总之，rnai介导的uPAR和组织蛋白酶B的下调启动了部分外源性凋亡级联，并伴有AIF的核易位。
The invasive character of gliomas depends on proteolytic cleavage of the surrounding extracellular matrix. Cathepsin B and urokinase-type plasminogen activator receptor (uPAR) together are known to be overexpressed in gliomas and, as such, are attractive targets for gene therapy. In the present study, we used plasmid constructs to induce the RNA interference (RNAi)-mediated down-regulation of uPAR and cathepsin B in SNB19 human glioma cells. We observed that the simultaneous down-regulation of uPAR and cathepsin B induces the up-regulation of proapoptotic genes and initiates a collapse in mitochondrial Delta psi. Cathepsin B and uPAR down-regulated cells showed increases in the expression of activated caspase-8 and DFF40/caspase-activated DNase. Nuclear translocation of AIF and Fas ligand translocation to the cell membrane were also observed. Ki67 and X-linked inhibitor of apoptosis protein levels decreased, thereby indicating apoptosis. These results suggest the involvement of uPAR-cathepsin B complex on the cell surface and its role in maintaining the viability of SNB19 glioma cells. In conclusion, RNAi-mediated down-regulation of uPAR and cathepsin B initiates a partial extrinsic apoptotic cascade accompanied by the nuclear translocation of AIF.