长链非编码RNA NORAD通过吸附miR-136-5p/PBX3轴促进视网膜母细胞瘤进展。

Long noncoding RNA NORAD promotes the progression of retinoblastoma by sponging miR-136-5p/PBX3 axis.

作者信息

Yang X-L, Hao Y-J, Wang B, Gu X-L, Wang X-X, Sun J-F

机构信息

Department of Ophthalmology, Qilu Hospital of Shandong University, Jinan, Shandong Province, China.

出版信息

Eur Rev Med Pharmacol Sci. 2020 Jan;24(3):1278-1287. doi: 10.26355/eurrev_202001_20185.

DOI:10.26355/eurrev_202001_20185

PMID:32096159

Abstract

OBJECTIVE

The specific roles of long noncoding RNAs (lncRNAs) have been found in human cancers, including retinoblastoma (RB). However, the function of lncRNA-NORAD has not been reported in RB. Therefore, the regulatory mechanism of lncRNA-NORAD was investigated in the development of RB.

PATIENTS AND METHODS

The experimental tissues were collected from 24 RB patients and 6 patients with ruptured globes. The average age of all patients was 2.78 years (range, 2 months to 11 years). The mRNA and protein expression was measured by Real Time-quantitative Polymerase Chain Reaction (RT-qPCR) and Western blot analysis. The functional mechanism of NORAD was assessed by Cell Counting Kit-8 (CCK-8), transwell, and Dual-Luciferase reporter assays.

RESULTS

Upregulation of NORAD and downregulation of miR-136-5p were found in RB. Functionally, knockdown of NORAD and miR-136-5p overexpression restrained RB cell viability, invasion, and migration. In addition, NORAD acts as a ceRNA of miR-136-5p in RB. MiR-136-5p was found to directly target PBX3. Furthermore, knockdown of PBX3 inhibited the progression of RB. More importantly, the NORAD/miR-136-5p axis is involved in RB progression by mediating PBX3.

CONCLUSIONS

LncRNA NORAD, serving as a ceRNA of miR-136-5p, accelerates RB progression by upregulation of PBX3.

摘要

目的

长链非编码RNA（lncRNA）在包括视网膜母细胞瘤（RB）在内的人类癌症中发挥了特定作用。然而，lncRNA-NORAD在RB中的功能尚未见报道。因此，本研究探讨了lncRNA-NORAD在RB发生发展中的调控机制。

患者与方法

收集24例RB患者及6例眼球破裂患者的实验组织。所有患者的平均年龄为2.78岁（范围为2个月至11岁）。通过实时定量聚合酶链反应（RT-qPCR）和蛋白质免疫印迹分析检测mRNA和蛋白质表达。通过细胞计数试剂盒-8（CCK-8）、Transwell和双荧光素酶报告基因检测评估NORAD的功能机制。

结果

RB中发现NORAD上调和miR-136-5p下调。在功能上，敲低NORAD和过表达miR-136-5p可抑制RB细胞的活力、侵袭和迁移。此外，NORAD在RB中作为miR-136-5p的竞争性内源性RNA（ceRNA）发挥作用。发现miR-136-5p直接靶向PBX3。此外，敲低PBX3可抑制RB的进展。更重要的是，NORAD/miR-136-5p轴通过介导PBX3参与RB的进展。