Targeted knockdown of Notch1 inhibits invasion of human prostate cancer cells concomitant with inhibition of matrix metalloproteinase-9 and urokinase plasminogen activator.
Targeted knockdown of Notch1 inhibits invasion of human prostate cancer cells concomitant with inhibition of matrix metalloproteinase-9 and urokinase plasminogen activator.
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DOI：
10.1158/1078-0432.ccr-08-1631
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发表时间：
2009-01-15
期刊：
影响因子：
--
通讯作者：
Mukhtar H
中科院分区：
文献类型：
--
作者：
Bin Hafeez B;Adhami VM;Asim M;Siddiqui IA;Bhat KM;Zhong W;Saleem M;Din M;Setaluri V;Mukhtar H
Notch是一种1型跨膜蛋白，在许多组织和器官类型的发育中起着关键作用。在多种人类癌症中发现的异常Notch信号传导有助于肿瘤的发展。由于发现Notch 1在前列腺癌（PCa）细胞和人PCa组织中过表达，因此我们测试了我们的假设，即PCa中Notch 1的过表达促进肿瘤侵袭。在人PCa细胞和人PCa组织中评估Notch 1表达。用浓度范围为30-120 nM的Notch 1特异性siRNA瞬时转染PCa细胞，随后评估对侵袭的影响和参与侵袭的分子的表达分析。小干扰RNA介导的PCa PC 3和22 R ν1细胞中Notch 1的敲低显著降低了它们的侵袭性。聚焦cDNA阵列显示Notch 1基因敲低导致尿激酶纤溶酶原激活物（uPA）和基质金属蛋白酶（MMP）-9基因转录本的表达显著降低。RT-PCR、真实的实时RT-PCR和免疫印迹分析进一步验证了上述结果。Notch 1的敲除也可显著降低uPA及其受体uPAR的mRNA表达和蛋白水平。Notch 1敲除细胞中MMP 9表达的显著降低表明Notch 1在增强MMP 9转录中的作用。我们的数据表明Notch 1参与人前列腺癌的侵袭，Notch 1的沉默通过抑制MMP 9和uPA的表达来抑制人前列腺癌细胞的侵袭。因此，靶向Notch 1可能是针对PCa的有效治疗方法。
Notch, a type 1 transmembrane protein, plays a key role in the development of many tissues and organ types. Aberrant Notch signaling found in a wide variety of human cancers contributes to tumor development. Since Notch1 was found to be over-expressed in prostate cancer (PCa) cells, and human PCa tissue we, therefore tested our hypothesis that over-expression of Notch1 in PCa promotes tumor invasion. Notch1 expression was evaluated in human PCa cells and in human PCa tissues. PCa cells were transiently transfected with Notch1 specific siRNAs in concentrations ranging from 30-120 nM and subsequently evaluated for effects on invasion and expression analysis for molecules involved in invasion. Small interfering RNA mediated knockdown of Notch1 in PCa PC3 and 22Rν1 cells dramatically decreased their invasion. Focused cDNA array revealed that Notch1 knockdown resulted in significant reduction in the expression of urokinase plasminogen activator (uPA) and matrix metalloproteinase (MMP)-9 gene transcripts. These data were further verified by RT-PCR, real- time RT-PCR and immunoblot analysis. Knockdown of Notch1 was also observed to significantly reduce the mRNA expression and protein levels of uPA and its receptor uPAR. A significant reduction in MMP9 expression in Notch1 knockdown cells suggested a role for Notch1 in augmenting MMP9 transcription. Our data demonstrate involvement of Notch1 in human PCa invasion and that silencing of Notch1 inhibits invasion of human PCa cells by inhibiting the expression of MMP9 and uPA. Thus, targeting of Notch1 could be an effective therapeutic approach against PCa.