Mao Wenjun, Wang Shengfei, Chen Ruo, He Yijun, Lu Rongguo, Zheng Mingfeng
Department of Cardiothoracic Surgery, The Affiliated Wuxi People's Hospital of Nanjing Medical University, Wuxi, 214023, Jiangsu, China.
Department of Cardiothoracic Surgery, The Affiliated Wuxi People's Hospital of Nanjing Medical University, No. 299 Qingyang Road, Wuxi, 214023, Jiangsu, China.
Open Med (Wars). 2022 Sep 28;17(1):1538-1549. doi: 10.1515/med-2022-0538. eCollection 2022.
Lung cancer (LC) is a prevailing primary tumor in the lung. lncRNA non-coding RNA activated by DNA damage (NORAD) is a popular target in human cancers. This experiment is designed to probe the mechanism of lncRNA in LC progression. NORAD expression in normal lung epithelial cells and LC cells was examined and then silenced to assess its effect on LC cell proliferation, invasion, and migration. Subcellular localization of NORAD was analyzed through online databases and then corroborated by fractionation of nuclear and cytoplasmic RNA assay. The target binding relations between NORAD and miR-28-3p and between miR-28-3p and E2F2 were verified. Eventually, LC cells with NORAD silencing were transfected with miR-28-3p inhibitor or pcDNA3.1-E2F2 to measure LC cell proliferation, invasion, and migration. NORAD was overexpressed in LC cells and NORAD knockout led to suppressed LC cell proliferation, invasion, and migration. Besides, NORAD targeted miR-28-3p and miR-28-3p targeted E2F2 transcription. Inhibiting miR-28-3p or overexpressing E2F2 could both annul the inhibitory role of si-NORAD in LC cell proliferation, invasion, and migration. Generally, our findings demonstrated that NORAD competitively bound to miR-28-3p with E2F2, to promote LC cell progression.
肺癌（LC）是肺部常见的原发性肿瘤。DNA损伤激活的长链非编码RNA（lncRNA）（NORAD）是人类癌症中一个热门的研究靶点。本实验旨在探究lncRNA在肺癌进展中的作用机制。检测了正常肺上皮细胞和肺癌细胞中NORAD的表达，然后将其沉默以评估其对肺癌细胞增殖、侵袭和迁移的影响。通过在线数据库分析NORAD的亚细胞定位，然后通过核质RNA分离实验进行验证。验证了NORAD与miR-28-3p以及miR-28-3p与E2F2之间的靶向结合关系。最后，用miR-28-3p抑制剂或pcDNA3.1-E2F2转染沉默NORAD的肺癌细胞，以检测肺癌细胞的增殖、侵袭和迁移情况。NORAD在肺癌细胞中高表达，敲除NORAD可导致肺癌细胞增殖、侵袭和迁移受到抑制。此外，NORAD靶向miR-28-3p，miR-28-3p靶向E2F2转录。抑制miR-28-3p或过表达E2F2均可消除si-NORAD对肺癌细胞增殖、侵袭和迁移的抑制作用。总的来说，我们的研究结果表明，NORAD与E2F2竞争性结合miR-28-3p，从而促进肺癌细胞进展。
