Li Jun, Xu Xia, Wei Cungang, Liu Lei, Wang Tengqi
Department of Thoracic Surgery, Bayannaoer City Hospital Inner Mongolia, China.
Department of Gynaecology and Obstetrics, Dengkou County People Hospital Inner Mongolia, China.
Int J Clin Exp Pathol. 2020 Jan 1;13(1):1-13. eCollection 2020.
Lung cancer is one of the most common human cancers. Long noncoding RNA-activated by DNA damage (NORAD) is often upregulated and promotes cell progression in various human cancers; however, its function and possible mechanism in lung cancer remain largely unknown.
The expression levels of NORAD, miR-30a-5p and a disintegrin and metalloproteinase 19 (ADAM19) were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR). 3-(4, 5)-dimethylthiazole-2-y1)-2, 5-biphenyl tetrazolium bromide (MTT) assay, flow cytometry, and transwell assay were employed to detect cell proliferation, apoptosis, migration, and invasion abilities, respectively. Western blot was used to detect the protein expression of ADAM19. The interaction between miR-30a-5p and NORAD or ADAM19 was predicted by online software and confirmed by the dual-luciferase reporter assay.
The expression levels of NORAD and ADAM19 were increased and the expression level of miR-30a-5p was decreased in lung cancer tissues and cells. Knockdown of NORAD could inhibit cell proliferation, migration and invasion but promote apoptosis in lung cancer cells. In addition, NORAD directly interacted with miR-30a-5p and its overexpression reversed the anti-cancer role of miR-30a-5p in lung cancer. Moreover, miR-30a-5p directly targeted ADAM19 and its inhibition attenuated the inhibitory effect of ADAM19 knockdown on progression of lung cancer cells. Furthermore, NORAD functioned as a competing endogenous RNA (ceRNA) through sponging miR-30a-5p to regulate ADAM19 expression.
NORAD knockdown suppressed cell proliferation, migration and invasion but promoted cell apoptosis in lung cancer cells by regulating miR-30a-5p/ADAM19, providing a possible therapeutic strategy for lung cancer patients.
肺癌是人类最常见的癌症之一。DNA损伤激活的长链非编码RNA（NORAD）在多种人类癌症中常上调并促进细胞进展；然而，其在肺癌中的功能及可能机制仍知之甚少。
通过定量实时聚合酶链反应（qRT-PCR）评估NORAD、miR-30a-5p和去整合素金属蛋白酶19（ADAM19）的表达水平。分别采用3-(4,5)-二甲基噻唑-2-基)-2,5-二苯基溴化四氮唑（MTT）法、流式细胞术和Transwell法检测细胞增殖、凋亡、迁移和侵袭能力。蛋白质印迹法用于检测ADAM19的蛋白表达。通过在线软件预测miR-30a-5p与NORAD或ADAM19之间的相互作用，并通过双荧光素酶报告基因实验进行验证。
肺癌组织和细胞中NORAD和ADAM19的表达水平升高，miR-30a-5p的表达水平降低。敲低NORAD可抑制肺癌细胞的增殖、迁移和侵袭，但促进其凋亡。此外，NORAD直接与miR-30a-5p相互作用，其过表达逆转了miR-30a-5p在肺癌中的抗癌作用。而且，miR-30a-5p直接靶向ADAM19，其抑制减弱了ADAM19敲低对肺癌细胞进展的抑制作用。此外，NORAD通过海绵吸附miR-30a-5p作为竞争性内源RNA（ceRNA）来调节ADAM19的表达。
敲低NORAD通过调节miR-30a-5p/ADAM19抑制肺癌细胞的增殖、迁移和侵袭，但促进细胞凋亡，为肺癌患者提供了一种可能的治疗策略。
