Institute of Pediatric Research, Children's Hospital of Soochow University, Suzhou, Jiangsu, China.
Eur Rev Med Pharmacol Sci. 2020 Dec;24(24):12748-12756. doi: 10.26355/eurrev_202012_24174.
Long noncoding RNA (lncRNA) plays a vital role in the progression of various cancers. However, the potential mechanisms of NR2F1-AS1 in the tumorigenesis of neuroblastoma (NB) have not been determined.
The expression levels of NR2F1-AS1, miR-493 and TRIM2 were detected by RT-qPCR. The downstream target genes of NR2F1-AS1 or miR-493 were predicted by bioinformatics analysis (http://starbase.sysu.edu.cn/), which was further indicated by Luciferase reporter and RNA immunoprecipitation (RIP) assays. CCK-8, transwell, and TUNEL assays were performed to determine the viability, migration, invasion and apoptosis of NB cells.
NR2F1-AS1 was highly expressed and miR-493 was lowly expressed in NB tissues and cell lines. The high expression of NR2F1-AS1 was associated with poor prognosis in NB. NR2F1-AS1 knockdown inhibited proliferation, migration, and invasion, and accelerated apoptosis of NB cells. MiR-493 was a downstream target of NR2F1-AS1, and the silencing of miR-493 reversed NR2F1-AS1 knockdown-attenuated progression of NB. Moreover, TRIM2 was demonstrated to be directly targeted by miR-493, and the upregulation of TRIM2 could abolish the inhibitory effect of miR-493 overexpression on the progression of NB. Finally, it was found that NR2F1-AS1 regulated TRIM2 expression by sponging miR-493.
The present study demonstrated that NR2F1-AS1 promoted the progression of NB through the miR-493/TRIM2 axis. This finding may provide new insight into the treatment of NB.
长链非编码 RNA(lncRNA)在多种癌症的进展中发挥着重要作用。然而,NR2F1-AS1 在神经母细胞瘤(NB)发生中的潜在机制尚未确定。
通过 RT-qPCR 检测 NR2F1-AS1、miR-493 和 TRIM2 的表达水平。通过生物信息学分析(http://starbase.sysu.edu.cn/)预测 NR2F1-AS1 或 miR-493 的下游靶基因,进一步通过荧光素酶报告和 RNA 免疫沉淀(RIP)实验进行验证。CCK-8、Transwell 和 TUNEL 实验用于检测 NB 细胞的活力、迁移、侵袭和凋亡。
NR2F1-AS1 在 NB 组织和细胞系中高表达,miR-493 低表达。NR2F1-AS1 的高表达与 NB 的不良预后相关。NR2F1-AS1 敲低抑制 NB 细胞的增殖、迁移和侵袭,并促进细胞凋亡。miR-493 是 NR2F1-AS1 的下游靶基因,miR-493 的沉默逆转了 NR2F1-AS1 敲低减弱的 NB 进展。此外,TRIM2 被证明是 miR-493 的直接靶基因,上调 TRIM2 可以消除 miR-493 过表达对 NB 进展的抑制作用。最后,研究发现 NR2F1-AS1 通过海绵 miR-493 来调节 TRIM2 的表达。
本研究表明,NR2F1-AS1 通过 miR-493/TRIM2 轴促进 NB 的进展。这一发现可能为 NB 的治疗提供新的思路。