长链非编码RNA LINC01296对神经母细胞瘤自发消退的影响及其机制研究

Although INSS stage 4S neuroblastomas (NBs) are metastatic, they are prone to undergo spontaneous regression and have a very good prognosis. The relationship between long non-coding RNAs (lncRNAs) and spontaneous regression of NB is yet to be investigated. We screened out lncRNA LINC01296 as a potential important lncRNA involved in NB spontaneous regression through mining gene microarrays data. Based on the results of bioinformatics analysis and primary experiments, we propose a hypothesis: LINC01296 infects the subcellular location of FMRP/BC200 and down-regulates the ERK & JNK signaling pathway in NB through direct interaction with FMRP/BC200, which result in inhibition of the apoptosis and differentiation of NB cells. In this project, we will probe the direct interaction among LINC01296, FMRP and BC200, detect the influence of LINC01296 on the subcellular location of FMRP and BC200, explore their regulation of ERK and JNK signaling pathway, investigate their impact on the tumor biological behavior of NB cells, and assess their clinical significance with the results of clinical tissue samples detection. This project is aimed at revealing the the influence of LINC01296 on spontaneous regression of NB as well as its underlying molecular mechanism, which promises in providing new theoretical foundation and therapeutic target for the treatment of progressive NB.

长链非编码RNA（lncRNA）与神经母细胞瘤（NB）自发消退的关系尚不明确。申请者通过基因芯片筛选出lncRNA LINC01296影响NB自发消退。结合前期研究，申请者提出以下假说：LINC01296通过与FMRP/BC200相互作用，改变后二者核/浆定位并下调ERK、JNK信号通路，从而抑制NB细胞凋亡及分化。本项目将围绕LINC01296-FMRP-BC200三者间的互作调控模式在NB中继续深入研究：明确LINC01296-FMRP-BC200三者间的相互作用；探明LINC01296对FMRP、BC200核/浆定位的影响；阐明三者互作对ERK、JNK信号通路活性的影响；细胞及裸鼠实验全面分析LINC01296对NB肿瘤生物学行为的影响；同时临床标本检测明确其临床意义。本项目旨在揭示LINC01296对NB自发消退的影响及分子机制，有望为治疗进展期NB提供新的理论依据和治疗靶点。

神经母细胞瘤（Neuroblastoma，NB）是最常见的儿童颅外恶性实体肿瘤。临床上，4期和4S期NB均已发生远处转移。但不同于4期NB，4S期NB具有非常高的自发消退比例。通过对4期及4S期NB组织标本基因表达谱芯片进行差异分析，我们发现长链非编码RNA LINC01296在4期NB中高表达，而在4S期NB中低表达。这提示LINC01296可能影响了NB的自发消退过程。细胞实验和动物实验表明，LINC01296可以显著促进NB细胞的增殖和侵袭能力，而抑制其凋亡和分化倾向。我们同时发现，LINC01296可以和FMRP蛋白相互结合，并促进FMRP蛋白在细胞核中的表达，但是FMRP进入细胞核中后是如何发挥作用的还不太清楚。我们也发现，LINC01296可以通过与FMRP蛋白相互作用，抑制NB细胞中MAPK信号通路的活性。对NB组织标本基因芯片分析显示，LINC01296与神经母细胞瘤患者的临床特征显著相关，死亡组患者LINC01296的表达水平显著高于存活组患者，而生存曲线显示高表达LINC01296的患者总体生存率及无事件生存率都显著要差于低表达LINC01296的患者。另外，RNA测序显示LINC01296的下游靶基因与免疫反应及神经发育密切相关，这些需要进一步进行挖掘研究。