RNAi-mediated downregulation of urokinase plasminogen activator receptor and matrix metalloprotease-9 in human breast cancer cells results in decreased tumor invasion, angiogenesis and growth (Retracted Article)
RNAi-mediated downregulation of urokinase plasminogen activator receptor and matrix metalloprotease-9 in human breast cancer cells results in decreased tumor invasion, angiogenesis and growth (Retracted Article)
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DOI：
10.1002/ijc.22962
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发表时间：
2007-11-15
影响因子：
6.4
通讯作者：
Rao, Jasti S.
中科院分区：
文献类型：
--
作者：
Kunigal, Sateesh;Lakka, Sajani S.;Rao, Jasti S.
丝氨酸蛋白酶尿激酶型纤溶酶原激活剂（uPA）在与其特异性受体uPAR（也称为CD 87）结合时在肿瘤细胞侵袭和转移中起重要作用。除了uPA-uPAR系统外，基质金属蛋白酶（MMPs）参与肿瘤细胞的侵袭和转移。本研究利用RNAi技术实现了对uPAR和MMP-9的特异性抑制。我们用小分子干扰RNA（siRNA）下调乳腺癌细胞系MDA MB 231和ZR 75 1中uPAR和MMP-9（pUM）的表达。在体外血管生成的研究表明，在处理的细胞的血管生成的潜力下降，特别是，一个显着的减少，观察到在与双顺反子构建体（PUM）处理的细胞相比，对照组。此外，双顺反子构建体抑制血管生成的体内模型中的毛细血管样结构的形成。类似地，如基质胶侵袭和迁移测定所示，当用双顺反子构建体处理时，侵袭潜力和迁移显著降低。这些结果表明uPAR和MMP-9的同时下调具有协同效应。我们还评估了MAPK、ERK和AKT信号通路分子的磷酸化形式的水平，发现与对照细胞相比，用双顺反子构建体处理的细胞中这些分子的水平降低。此外，靶向uPAR和MMP-9完全消退裸鼠原位乳腺肿瘤。总之，我们的研究结果提供了证据表明，使用RNAi技术同时下调uPAR和MMP-9可能为乳腺癌治疗提供一种有效的工具。
The serine protease urokinase-type plasminogen activator (uPA) plays a significant role in tumor cell invasion and metastasis when bound to its specific receptor, uPAR (also known as CD87). In addition to the uPA-uPAR system, matrix metalloproteinases (MMPs) are involved in tumor cell invasion and metastasis. In this study, we achieved specific inhibition of uPAR and MMP-9 using RNAi technology. We introduced small interfering RNA to down-regulate the expression of uPAR and MMP-9 (pUM) in breast cancer cell lines (MDA MB 231 and ZR 75 1). In vitro angiogenesis studies indicated a decrease in the angiogenic potential of the treated cells; in particular, a remarkable decrease was observed in the cells treated with bicistronic construct (pUM) in comparision to the controls. Additionally, bicistronic construct inhibited the formation of capillary-like structures in in vivo models of angiogenesis. Similarly, the invasive potential and migration decreased dramatically when treated with the bicistronic construct as shown by matrigel invasion and migration assays. These results suggest a synergistic effect from the simultaneous downregulation of uPAR and MMP-9. We also assessed the levels of phosphorylated forms of MAPK, ERK and AKT signaling pathway molecules and found reduction in the levels of these molecules in cells treated with the bicistronic construct as compared to the control cells. Furthermore, targeting both uPAR and MMP-9 totally regressed orthotopic breast tumors in nude mice. In conclusion, our results provide evidence that the simultaneous downregulation of uPAR and MMP-9 using RNAi technology may provide an effective tool for breast cancer therapy.