Qiao Jingbo, Kang Jung-Hee, Cree Jeremy, Evers B Mark, Chung Dai H
Department of Surgery, The University of Texas Medical Branch, Galveston, TX 77555-0353, USA.
Neoplasia. 2007 Mar;9(3):184-91. doi: 10.1593/neo.06841.
Angiogenesis plays a critical role in tumor progression in various cancers, including neuroblastoma. We have previously shown that gastrin-releasing peptide (GRP) stimulates neuroblastoma growth and that its cell surface receptors, gastrin-releasing peptide receptors (GRP-R), are overexpressed in advanced-stage human neuroblastomas; however, the effects of GRP on angiogenesis are not clearly elucidated. Interleukin (IL) 8, a proinflammatory chemokine, plays an important role during tumor angiogenesis. Ets transcription factors, such as oncoproteins, cause tumor development and are also known to induce IL-8 expression. In the present study, we found an increased expression of Ets1 in more undifferentiated human neuroblastomas. Stable transfection of SK-N-SH human neuroblastoma cells with Ets1 plasmid resulted in increased IL-8 luciferase activity and IL-8 secretion into cell culture media. Conversely, silencing of Ets1 resulted in a significant decrease in IL-8 secretion in SK-N-SH cells. Moreover, exogenous GRP treatment increased Ets1 (T38) phosphorylation and Ets1 nuclear accumulation, and enhanced Ets1 binding to its DNA consensus sequence, resulting in the stimulation of IL-8 mRNA expression and protein secretion. Our findings demonstrate that GRP upregulates proangiogenic IL-8 expression in an Ets1-dependent manner, suggesting a critical role of this process during GRP-induced neuroblastoma angiogenesis and metastasis.
血管生成在包括神经母细胞瘤在内的多种癌症的肿瘤进展中起着关键作用。我们之前已经表明,胃泌素释放肽(GRP)刺激神经母细胞瘤生长,并且其细胞表面受体胃泌素释放肽受体(GRP-R)在晚期人类神经母细胞瘤中过度表达;然而,GRP对血管生成的影响尚未明确阐明。白细胞介素(IL)8是一种促炎趋化因子,在肿瘤血管生成过程中起重要作用。Ets转录因子,如癌蛋白,可导致肿瘤发展,并且也已知可诱导IL-8表达。在本研究中,我们发现Ets1在更多未分化的人类神经母细胞瘤中表达增加。用Ets1质粒稳定转染SK-N-SH人神经母细胞瘤细胞导致IL-8荧光素酶活性增加以及IL-8分泌到细胞培养基中。相反,Ets1沉默导致SK-N-SH细胞中IL-8分泌显著减少。此外,外源性GRP处理增加了Ets1(T38)磷酸化和Ets1核积累,并增强了Ets1与其DNA共有序列的结合,导致IL-8 mRNA表达和蛋白质分泌受到刺激。我们的研究结果表明,GRP以Ets1依赖性方式上调促血管生成的IL-8表达,表明该过程在GRP诱导的神经母细胞瘤血管生成和转移中起关键作用。