Center for Clinical Molecular Medicine, National Clinical Research Center for Child Health and Disorders, Ministry of Education Key Laboratory of Child Development and Disorders, China International Science and Technology Cooperation Base of Child Development and Critical Disorders, Chongqing Key Laboratory of Pediatrics, Children's Hospital of Chongqing Medical University, Chongqing, 400014, China.
Chongqing Key Laboratory of Translational Research for Cancer Metastasis and Individualized Treatment, Chongqing University Cancer Hospital, Chongqing, 400030, China.
J Transl Med. 2023 Sep 23;21(1):659. doi: 10.1186/s12967-023-04535-3.
Oncogene MYCN is closely related with malignant progression and poor prognosis of neuroblastoma (NB). Recently, long non-coding RNAs (lncRNAs) have been recognized as crucial regulators in various cancers. However, whether lncRNAs contribute to the overexpression of MYCN in NB is unclear.
Microarray analysis were applied to analyze the differentially expressed lncRNAs between MYCN-amplified and MYCN-non-amplified NB cell lines. Bioinformatic analyses were utilized to identify lncRNAs nearby MYCN locus. qRT-PCR was used to detect the expression level of lncRNA AC142119.1 in NB cell lines and tissues. Gain- and loss-of-function assays were conducted to investigate the biological effect of AC142119.1 in NB. Fluorescence in situ hybridization, RNA pull-down, RNA immunoprecipitation, mass spectrometry, RNA electrophoretic mobility shift, chromatin immunoprecipitation and chromatin isolation by RNA purification assays were performed to validate the interaction between AC142119.1 and WDR5 protein as well as MYCN promoter.
AC142119.1 was significantly elevated in NB tissues with MYCN amplification, advanced INSS stage and high risk, and associated with poor survival of NB patients. Moreover, enforced expression of AC142119.1 reinforced the proliferation of NB cells in vitro and in vivo. Additionally, AC142119.1 specifically recruited WDR5 protein to interact with MYCN promoter, further initiating the transcription of MYCN and accelerating NB progression.
We identified a novel lncRNA AC142119.1, which promoted the progression of NB through epigenetically initiating the transcription of MYCN via interacting with both WDR5 protein and the promoter of MYCN, indicating that AC142119.1 might be a potential diagnostic biomarker and therapeutic target for NB.
癌基因 MYCN 与神经母细胞瘤（NB）的恶性进展和不良预后密切相关。最近，长链非编码 RNA（lncRNA）已被认为是各种癌症中重要的调控因子。然而，lncRNA 是否导致 NB 中 MYCN 的过度表达尚不清楚。
应用微阵列分析方法分析 MYCN 扩增和非扩增 NB 细胞系之间差异表达的 lncRNA。生物信息学分析用于鉴定 MYCN 基因座附近的 lncRNA。qRT-PCR 用于检测 NB 细胞系和组织中 lncRNA AC142119.1 的表达水平。通过 gain-和 loss-of-function 实验研究 AC142119.1 在 NB 中的生物学效应。荧光原位杂交、RNA 下拉、RNA 免疫沉淀、质谱、RNA 电泳迁移率变动、染色质免疫沉淀和 RNA 纯化染色质分离实验用于验证 AC142119.1 与 WDR5 蛋白以及 MYCN 启动子之间的相互作用。
AC142119.1 在 MYCN 扩增、INSS 晚期和高危的 NB 组织中显著升高，与 NB 患者的不良生存相关。此外，AC142119.1 的强制表达增强了 NB 细胞在体外和体内的增殖能力。此外，AC142119.1 特异性募集 WDR5 蛋白与 MYCN 启动子相互作用，进而启动 MYCN 的转录，加速 NB 的进展。
我们鉴定了一种新型 lncRNA AC142119.1，通过与 WDR5 蛋白和 MYCN 启动子相互作用，表观遗传地启动 MYCN 的转录，从而促进 NB 的进展，表明 AC142119.1 可能是 NB 的潜在诊断生物标志物和治疗靶点。
