Effects of a Novel Long Noncoding RNA, IncUSMycN, on N-Myc Expression and Neuroblastoma Progression
Effects of a Novel Long Noncoding RNA, IncUSMycN, on N-Myc Expression and Neuroblastoma Progression
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DOI：
10.1093/jnci/dju113
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发表时间：
2014-07-01
影响因子：
10.3
通讯作者：
Liu, Tao
中科院分区：
文献类型：
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作者：
Liu, Pei Y.;Erriquez, Daniela;Liu, Tao
背景含有MYCN癌基因的130kb基因组DNA扩增导致神经母细胞瘤患者预后不良。方法利用生物信息学数据，在130kb扩增片段上发现一个新的非编码RNA IncUSMycN。RNA-蛋白质下拉试验用于鉴定与IncUSMycN RNA结合的蛋白质。采用Kaplan-Meier生存分析、多变量COX回归分析和双侧LOG检验，分析IncUSMycN和NONO在3组神经母细胞瘤患者(n=47、88和476)中的预后价值。用针对IncUSMycN的反义寡核苷酸(n=12)或错配序列(n=13)处理神经母细胞瘤荷瘤小鼠，结果进行多重比较双向方差分析。结果生物信息学数据预测了IncUSMycN基因和RNA，逆转录聚合酶链式反应证实了其3个外显子和2个内含子。在341例神经母细胞瘤组织中，88例与MYCN共扩增出IncUSMycN基因。IncUSMycN RNA结合RNA结合蛋白NONO，导致N-Myc RNA上调和神经母细胞瘤细胞增殖。神经母细胞瘤组织中IncUSMycN和NONO的高表达独立预测患者预后不良(INcUSMycN：危险比[HR]=1.87，95%可信区间[CI]=1.06~3.28，P=0.03；NONO：HR=2.48，95%CI=1.34~4.57，P=.004)。在神经母细胞瘤荷瘤小鼠中，针对IncUSMycN的反义寡核苷酸治疗在统计学上显著地阻止了肿瘤的进展(P<；.001)。结论IncUSMycN和NONO在调节N-Myc表达和神经母细胞瘤的发生中发挥了重要作用，并首次提供了扩增长非编码RNA基因有助于肿瘤发生的证据。
Background Patients with neuroblastoma due to the amplification of a 130-kb genomic DNA region containing the MYCN oncogene have poor prognoses.Methods Bioinformatics data were used to discover a novel long noncoding RNA, IncUSMycN, at the 130-kb amplicon. RNA-protein pull-down assays were used to identify proteins bound to IncUSMycN RNA. Kaplan-Meier survival analysis, multivariable Cox regression, and two-sided log-rank test were used to examine the prognostic value of IncUSMycN and NonO expression in three cohorts of neuroblastoma patients (n = 47, 88, and 476, respectively). Neuroblastoma-bearing mice were treated with antisense oligonucleotides targeting IncUSMycN (n = 12) or mismatch sequence (n = 13), and results were analyzed by multiple comparison two-way analysis of variance. All statistical tests were two-sided.Results Bioinformatics data predicted IncUSMycN gene and RNA, and reverse-transcription polymerase chain reaction confirmed its three exons and two introns. The IncUSMycN gene was coamplified with MYCN in 88 of 341 human neuroblastoma tissues. IncUSMycN RNA bound to the RNA-binding protein NonO, leading to N-Myc RNA upregulation and neuroblastoma cell proliferation. High levels of IncUSMycN and NonO expression in human neuroblastoma tissues independently predicted poor patient prognoses (IncUSMycN: hazard ratio [HR] = 1.87, 95% confidence interval [CI] = 1.06 to 3.28, P = .03; NonO: HR = 2.48, 95% CI = 1.34 to 4.57, P = .004). Treatment with antisense oligonucleotides targeting IncUSMycN in neuroblastoma-bearing mice statistically significantly hindered tumor progression (P < .001).Conclusions Our data demonstrate the important roles of IncUSMycN and NonO in regulating N-Myc expression and neuroblastoma oncogenesis and provide the first evidence that amplification of long noncoding RNA genes can contribute to tumorigenesis.