Department of Respiratory Medicine, Jiangsu Taizhou People's Hospital, Taizhou, China.
Mol Genet Genomic Med. 2019 Aug;7(8):e757. doi: 10.1002/mgg3.757. Epub 2019 Jun 17.
To explore the role of non-coding RNA activated by DNA damage (NORAD), a long non-coding ribonucleic acid (lncRNA), in non-small cell lung cancer (NSCLC) and its possible mechanism.
Quantitative real-time polymerase chain reaction was adopted for the detection of the expression levels of NORAD, micro RNA (miR)-656-3p, and AKT serine/threonine kinase 1 (AKT1). The effects of NORAD, miR-656-3p, and AKT1 on cell proliferation and migration were examined through the Cell Counting Kit-8 (CCK-8) and Transwell assay. Subsequently, the binding relationships between miR-656-3p and AKT1 and between miR-656-3p and NORAD were verified by dual-luciferase reporter gene assay. Finally, the potential mechanisms of action of NORAD and miR-656-3p were explored through the torsion experiment.
The lncRNA NORAD expression level in NSCLC patients was notably higher than that in people in control group, that in patients with metastasis was higher than that in patients without metastasis, and that in patients with NSCLC in stage III-IV was significantly higher than that in patients with NSCLC in stage I-II. Elevation of NORAD stimulated the proliferation and migration of NSCLC A549/H460 cells. According to the reporter gene assay, NORAD could bind to miR-656-3p. Besides, miR-656-3p was significantly under-expressed in cancer tissues of patients with NSCLC, and overexpression of miR-656-3p could block the proliferation and migration of A549/H460 cells and reversed promotion on cell proliferation and migration by NORAD. Furthermore, the reporter gene assay revealed that the overexpression of AKT1, a miR-656-3p target gene, could reverse miR-656-3p's inhibitory effect on the proliferation and migration of A549/H460 cells.
LncRNA NORAD is capable of promoting the proliferation and migration of NSCLC cells, and its mechanism may be that it increases the AKT1 expression by adsorbing miR-656-3p.
探讨 DNA 损伤激活的非编码 RNA（NORAD），一种长链非编码核糖核酸（lncRNA），在非小细胞肺癌（NSCLC）中的作用及其可能的机制。
采用实时定量聚合酶链反应检测 NORAD、微小 RNA（miR）-656-3p 和 AKT 丝氨酸/苏氨酸激酶 1（AKT1）的表达水平。通过细胞计数试剂盒-8（CCK-8）和 Transwell 实验检测 NORAD、miR-656-3p 和 AKT1 对细胞增殖和迁移的影响。随后，通过双荧光素酶报告基因实验验证 miR-656-3p 与 AKT1 以及 miR-656-3p 与 NORAD 之间的结合关系。最后，通过扭转实验探讨 NORAD 和 miR-656-3p 的潜在作用机制。
与对照组相比，NSCLC 患者的 lncRNA NORAD 表达水平明显升高，转移患者的表达水平高于无转移患者，III-IV 期 NSCLC 患者的表达水平显著高于 I-II 期患者。NORAD 的升高刺激了 NSCLC A549/H460 细胞的增殖和迁移。根据报告基因实验，NORAD 可以与 miR-656-3p 结合。此外，miR-656-3p 在 NSCLC 患者的癌组织中表达明显下调，过表达 miR-656-3p 可阻断 A549/H460 细胞的增殖和迁移，并逆转 NORAD 对细胞增殖和迁移的促进作用。此外，报告基因实验表明，miR-656-3p 靶基因 AKT1 的过表达可逆转 miR-656-3p 对 A549/H460 细胞增殖和迁移的抑制作用。
lncRNA NORAD 能够促进 NSCLC 细胞的增殖和迁移，其机制可能是通过吸附 miR-656-3p 增加 AKT1 的表达。
