Jordà Mireia, Olmeda David, Vinyals Antònia, Valero Eva, Cubillo Eva, Llorens Ana, Cano Amparo, Fabra Angels
Centre d'Oncologia Molecular, IDIBELL-Institut de Recerca Oncològica, Barcelona, Spain.
J Cell Sci. 2005 Aug 1;118(Pt 15):3371-85. doi: 10.1242/jcs.02465.
Overexpression of the transcription factor Snail in epithelial MDCK cells promotes the epithelial-mesenchymal transition (EMT) and the acquisition of an invasive phenotype. We report here that the expression of Snail is associated with an increase in the promoter activity and expression of the matrix metalloproteinase MMP-9. The effect of Snail silencing on MMP-9 expression corroborates this finding. Induced transcription of MMP-9 by Snail is driven by a mechanism dependent on the MAPK and phosphoinositide 3-kinase (PI3K) signalling pathways. Although other regions of the promoter were required for a complete stimulation by Snail, a minimal fragment (nucleotides -97 to +114) produces a response following an increased phosphorylation of Sp-1 and either Sp-1 or Ets-1 binding to the GC-box elements contained in this region. The expression of a dominant negative form of MEK decreased these complexes. A moderate increase in the binding of the nuclear factor kappaB (NFkappaB) to the upstream region (nucleotide -562) of the MMP-9 promoter was also observed in Snail-expressing cells. Interestingly, oncogenic H-Ras (RasV12) synergistically co-operates with Snail in the induction of MMP-9 transcription and expression. Altogether, these results indicate that MMP-9 transcription is activated in response to Snail expression and that it might explain, at least in part, the invasive properties of the Snail-expressing cells.
转录因子Snail在上皮性MDCK细胞中的过表达促进上皮-间质转化(EMT)并获得侵袭性表型。我们在此报告,Snail的表达与基质金属蛋白酶MMP-9的启动子活性增加及表达上调相关。Snail沉默对MMP-9表达的影响证实了这一发现。Snail诱导的MMP-9转录是由一种依赖于丝裂原活化蛋白激酶(MAPK)和磷脂酰肌醇3-激酶(PI3K)信号通路的机制驱动的。尽管启动子的其他区域对于Snail的完全刺激是必需的,但一个最小片段(核苷酸-97至+114)在Sp-1磷酸化增加以及Sp-1或Ets-1与该区域所含GC盒元件结合后会产生反应。MEK显性负性形式的表达减少了这些复合物。在表达Snail的细胞中还观察到核因子κB(NFκB)与MMP-9启动子上游区域(核苷酸-562)的结合适度增加。有趣的是,致癌性H-Ras(RasV12)在诱导MMP-9转录和表达方面与Snail协同合作。总之,这些结果表明MMP-9转录因Snail表达而被激活,这可能至少部分解释了表达Snail的细胞的侵袭特性。