Hu Yanping, Sun Huifang, Hu Jiting, Zhang Xiaomin
Department of Pediatrics, Luoyang Central Hospital Affiliated to Zhengzhou University, Luoyang, Henan 471009, People's Republic of China.
Cancer Manag Res. 2020 Aug 19;12:7451-7463. doi: 10.2147/CMAR.S252521. eCollection 2020.
Neuroblastoma (NB) is a common malignant tumor of the sympathetic nervous system, mainly disturbing children. Long non-coding RNAs (lncRNAs) serving as promising cancer biomarkers have been well recognized. Our study intends to explore the functions of lncRNA X-inactive specific transcript (DLX6-AS1) in NB and provide a potential action mechanism.
The expression of DLX6-AS1, miR-506-3p and signal transducer and activator of transcription 2 () was measured by quantitative real-time polymerase chain reaction (qRT-PCR). Cell proliferation was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and colony formation assay. Cell cycle distribution was determined by flow cytometry assay. The protein level of cell cycle-related markers and STAT2 was detected by Western blot. Glycolysis progress was evaluated according to glucose consumption, lactate production and ATP level. The target genes were predicted by the online database Starbase3.0 and verified by dual-luciferase reporter assay.
DLX6-AS1 expression was highly elevated in NB tissues and cells. DLX6-AS1 deficiency inhibited NB cell proliferation, cell cycle and glycolysis in vitro. MiR-506-3p was a target of DLX6-AS1, and miR-506-3p absence partly reversed the effects of DLX6-AS1 deficiency. Besides, STAT2 was targeted by miR-506-3p, and its expression was regulated by DLX6-AS1 through miR-506-3p. MiR-506-3p restoration also inhibited NB cell malignant behaviors, and overexpression partially abolished the role of miR-506-3p restoration. Moreover, DLX6-AS1 deficiency weakened tumor growth in vivo.
DLX6-AS1 regulated cell proliferation, cell cycle and glycolysis in vitro and tumor growth in vivo to promote the development of NB by upregulating via targeting miR-506-3p.
神经母细胞瘤（NB）是一种常见的交感神经系统恶性肿瘤，主要影响儿童。长链非编码RNA（lncRNA）作为有前景的癌症生物标志物已得到广泛认可。本研究旨在探讨lncRNA X失活特异性转录本（DLX6-AS1）在NB中的作用，并提供潜在的作用机制。
采用定量实时聚合酶链反应（qRT-PCR）检测DLX6-AS1、miR-506-3p和信号转导与转录激活因子2（STAT2）的表达。使用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐（MTT）法和集落形成试验评估细胞增殖。通过流式细胞术检测细胞周期分布。采用蛋白质印迹法检测细胞周期相关标志物和STAT2的蛋白水平。根据葡萄糖消耗、乳酸产生和ATP水平评估糖酵解进程。通过在线数据库Starbase3.0预测靶基因，并通过双荧光素酶报告基因试验进行验证。
DLX6-AS1在NB组织和细胞中表达显著升高。DLX6-AS1缺失在体外抑制NB细胞增殖、细胞周期和糖酵解。miR-506-3p是DLX6-AS1的靶标，miR-506-3p缺失部分逆转了DLX6-AS1缺失的作用。此外，STAT2是miR-506-3p的靶标，其表达受DLX6-AS1通过miR-506-3p调控。miR-506-3p恢复也抑制NB细胞的恶性行为，STAT2过表达部分消除了miR-506-3p恢复的作用。此外，DLX6-AS1缺失减弱了体内肿瘤生长。
DLX6-AS1通过靶向miR-506-3p上调STAT2，在体外调节细胞增殖、细胞周期和糖酵解，在体内调节肿瘤生长，从而促进NB的发展。
