Regulation of matrix metalloproteinase genes by E2F transcription factors: Rb-Raf-1 interaction as a novel target for metastatic disease.
Regulation of matrix metalloproteinase genes by E2F transcription factors: Rb-Raf-1 interaction as a novel target for metastatic disease.
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DOI：
10.1158/0008-5472.can-11-2647
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发表时间：
2012-01-15
期刊：
影响因子：
11.2
通讯作者：
Chellappan SP
中科院分区：
文献类型：
--
作者：
Johnson JL;Pillai S;Pernazza D;Sebti SM;Lawrence NJ;Chellappan SP
Rb-E2F转录调控通路在细胞周期调控中发挥重要作用，但对其在侵袭和转移中的作用知之甚少。我们发现，许多参与癌细胞侵袭的基因，如基质金属蛋白酶，在它们的启动子上都有潜在的E2F结合位点。预测了所有23个人基质金属蛋白酶基因启动子上的E2F结合位点，其中许多含有多个E2F结合位点。研究表明，在非小细胞肺癌(NSCLC)中过度表达的MMP9、MMP14和MMP15等MMP9、MMP14和MMP15基因具有多个E2F结合位点，并受Rb-E2F途径的调控。染色质免疫沉淀分析表明，E2F1与MMP9、MMP14和MMP15启动子结合，瞬时转染实验表明这些启动子对E2F有应答作用。相应地，通过RNAi技术耗尽E2F家族成员减少了这些基因的表达，并相应地降低了胶原降解活性。此外，使用RB-Raf-1干扰物RRD-251通过抑制Raf-1与Rb的相互作用来激活Rb足以抑制MMP的转录。这导致了癌细胞在体外的侵袭和迁移减少，以及在小鼠尾静脉肺转移模型中转移灶的发展。这些结果提示，E2F转录因子可能通过调控基质金属蛋白酶基因而发挥促进转移的作用，靶向Rb-Raf-1相互作用是治疗转移性疾病的一种有前景的方法。
The Rb-E2F transcriptional regulatory pathway plays a major role in cell cycle regulation, but its role in invasion and metastasis is less understood. We find that many genes involved in the invasion of cancer cells, such as matrix metalloproteinases, have potential E2F binding sites in their promoters. E2F binding sites were predicted on all 23 human MMP gene promoters, many of which harbored multiple E2F binding sites. Studies presented here show that MMP genes such as MMP9, MMP14, and MMP15 which are overexpressed in non-small cell lung cancer (NSCLC) have multiple E2F binding sites and are regulated by the Rb-E2F pathway. Chromatin immunoprecipitation assays showed the association of E2F1 with the MMP9, MMP14, and MMP15 promoters and transient transfection experiments showed that these promoters are E2F responsive. Correspondingly, depletion of E2F family members by RNAi techniques reduced the expression of these genes with a corresponding reduction in collagen degradation activity. Further, activating Rb by inhibiting the interaction of Raf-1 with Rb using the Rb-Raf-1 disruptor RRD-251 was sufficient to inhibit MMP transcription. This led to reduced invasion and migration of cancer cells in vitro and metastatic foci development in a tail vein lung metastasis model in mice. These results suggest that E2F transcription factors may play a role in promoting metastasis through regulation of MMP genes, and that targeting the Rb-Raf-1 interaction is a promising approach for the treatment of metastatic disease.