Cell Biology and Physiology Division, CSIR-Indian Institute of Chemical Biology, Council of Scientific and Industrial Research, 4 Raja S. C. Mullick Road, Kolkata 700032, India.
J Biol Chem. 2012 Apr 27;287(18):15001-15. doi: 10.1074/jbc.M111.284034. Epub 2012 Jan 23.
Matrix metalloproteinase-mediated degradation of extracellular matrix is a crucial event for invasion and metastasis of malignant cells. The expressions of matrix metalloproteinases (MMPs) are regulated by different cytokines and growth factors. VEGF, a potent angiogenic cytokine, induces invasion of ovarian cancer cells through activation of MMPs. Here, we demonstrate that invasion and scattering in SKOV-3 cells were induced by VEGF through the activation of p38 MAPK and PI3K/AKT pathways. VEGF induced the expression of MMP-2, MMP-9, and MMP-13 and hence regulated the metastasis of SKOV-3 ovarian cancer cells, and the activities of these MMPs were reduced after inhibition of PI3K/AKT and p38 MAPK pathways. Interestingly, VEGF induced expression of ETS-1 factor, an important trans-regulator of different MMP genes. ETS-1 bound to both MMP-9 and MMP-13 promoters. Furthermore, VEGF acted through its receptor to perform the said functions. In addition, VEGF-induced MMP-9 and MMP-13 expression and in vitro cell invasion were significantly reduced after knockdown of ETS-1 gene. Again, VEGF-induced MMP-9 and MMP-13 promoter activities were down-regulated in ETS-1 siRNA-transfected cells. VEGF enriched ETS-1 in the nuclear fraction in a dose-dependent manner. VEGF-induced expression of ETS-1 and its nuclear localization were blocked by specific inhibitors of the PI3K and p38 MAPK pathways. Therefore, based on these observations, it is hypothesized that the activation of PI3K/AKT and p38 MAPK by VEGF results in ETS-1 gene expression, which activates MMP-9 and MMP-13, leading to the invasion and scattering of SKOV-3 cells. The study provides a mechanistic insight into the prometastatic functions of VEGF-induced expression of relevant MMPs.
细胞外基质的基质金属蛋白酶介导的降解是恶性细胞侵袭和转移的关键事件。基质金属蛋白酶(MMPs)的表达受不同细胞因子和生长因子的调节。VEGF 是一种有效的血管生成细胞因子,通过激活 MMPs 诱导卵巢癌细胞的侵袭。在这里,我们证明 VEGF 通过激活 p38 MAPK 和 PI3K/AKT 途径诱导 SKOV-3 细胞的侵袭和分散。VEGF 诱导 MMP-2、MMP-9 和 MMP-13 的表达,从而调节 SKOV-3 卵巢癌细胞的转移,并且这些 MMP 的活性在抑制 PI3K/AKT 和 p38 MAPK 途径后降低。有趣的是,VEGF 诱导 ETS-1 因子的表达,ETS-1 是不同 MMP 基因的重要转录调节剂。ETS-1 结合 MMP-9 和 MMP-13 启动子。此外,VEGF 通过其受体发挥上述功能。此外,在 ETS-1 基因敲低后,VEGF 诱导的 MMP-9 和 MMP-13 表达和体外细胞侵袭明显减少。同样,在转染 ETS-1 siRNA 的细胞中,VEGF 诱导的 MMP-9 和 MMP-13 启动子活性下调。VEGF 以剂量依赖的方式将 ETS-1 富集到核部分。PI3K 和 p38 MAPK 途径的特异性抑制剂阻断了 VEGF 诱导的 ETS-1 表达及其核定位。因此,基于这些观察结果,假设 VEGF 激活 PI3K/AKT 和 p38 MAPK 导致 ETS-1 基因表达,激活 MMP-9 和 MMP-13,导致 SKOV-3 细胞的侵袭和分散。该研究提供了关于 VEGF 诱导相关 MMP 表达的促转移功能的机制见解。