Planar cell polarity gene expression correlates with tumor cell viability and prognostic outcome in neuroblastoma.
Planar cell polarity gene expression correlates with tumor cell viability and prognostic outcome in neuroblastoma.
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DOI：
10.1186/s12885-016-2293-2
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发表时间：
2016-03-31
期刊：
影响因子：
3.8
通讯作者：
Johnsen JI
中科院分区：
文献类型：
--
作者：
Dyberg C;Papachristou P;Haug BH;Lagercrantz H;Kogner P;Ringstedt T;Wickström M;Johnsen JI
非经典Wnt/平面细胞极性（PCP）信号通路是胚胎发育过程中细胞迁移的主要参与者，最近被认为与肿瘤发生有关。使用cDNA质粒或siRNA转染来增加和抑制神经母细胞瘤细胞和非致瘤细胞中的Prickle 1和Vangl 2表达。通过台盼蓝拒染法测定细胞活力，并用蛋白质印迹法测定蛋白质表达。用荧光素酶报告基因检测转录活性，用实时荧光定量RT-PCR检测mRNA表达。免疫荧光染色用于研究Vangl 2过表达在非致瘤性胚胎细胞中的作用。统计学显著性用t检验或单因素方差分析检验。在这里，我们发现PCP核心基因Prickle 1和Vangl 2的高表达与低风险神经母细胞瘤、神经母细胞瘤细胞生长抑制和Wnt/β-连环蛋白信号转导减少相关。Rho相关激酶（ROCK）在介导非经典Wnt信号传导中起重要作用，抑制ROCK可导致神经母细胞瘤细胞中Prickle 1表达增加和β-连环蛋白活性抑制。相比之下，MYC永生化神经干细胞中Vangl 2的过表达诱导活性β-连环蛋白的积累并降低神经分化标志物Tujl。类似地，在nestin阳性细胞中强制过表达Vangl 2的遗传修饰小鼠在胚胎发育期间显示Tujl分化标记物减少。我们的实验数据表明，Prickle 1和Vangl 2的高表达减少了神经母细胞瘤细胞的生长，并表明PCP蛋白在致瘤细胞中与正常细胞相比的不同作用。这些结果表明，非经典的Wnt/PCP信号通路的活动是重要的神经母细胞瘤的发展和Wnt/PCP通路的操作提供了一种可能的治疗神经母细胞瘤。本文的在线版本（doi：10.1186/s12885-016-2293-2）包含补充材料，可供授权用户使用。
The non-canonical Wnt/Planar cell polarity (PCP) signaling pathway is a major player in cell migration during embryonal development and has recently been implicated in tumorigenesis. Transfections with cDNA plasmids or siRNA were used to increase and suppress Prickle1 and Vangl2 expression in neuroblastoma cells and in non-tumorigenic cells. Cell viability was measured by trypan blue exclusion and protein expression was determined with western blotting. Transcriptional activity was studied with luciferase reporter assay and mRNA expression with real-time RT-PCR. Immunofluorescence stainings were used to study the effects of Vangl2 overexpression in non-tumorigenic embryonic cells. Statistical significance was tested with t-test or one-way ANOVA. Here we show that high expression of the PCP core genes Prickle1 and Vangl2 is associated with low-risk neuroblastoma, suppression of neuroblastoma cell growth and decreased Wnt/β-catenin signaling. Inhibition of Rho-associated kinases (ROCKs) that are important in mediating non-canonical Wnt signaling resulted in increased expression of Prickle1 and inhibition of β-catenin activity in neuroblastoma cells. In contrast, overexpression of Vangl2 in MYC immortalized neural stem cells induced accumulation of active β-catenin and decreased the neural differentiation marker Tuj1. Similarly, genetically modified mice with forced overexpression of Vangl2 in nestin-positive cells showed decreased Tuj1 differentiation marker during embryonal development. Our experimental data demonstrate that high expression of Prickle1 and Vangl2 reduce the growth of neuroblastoma cells and indicate different roles of PCP proteins in tumorigenic cells compared to normal cells. These results suggest that the activity of the non-canonical Wnt/PCP signaling pathway is important for neuroblastoma development and that manipulation of the Wnt/PCP pathway provides a possible therapy for neuroblastoma. The online version of this article (doi:10.1186/s12885-016-2293-2) contains supplementary material, which is available to authorized users.