Cao Xiying, Gu Liang, Zhang Zuxiong, Zhong Weixiang, Guo Shaoming, Xie Chunfa
Cells Tissues Organs. 2022 Jun 14. doi: 10.1159/000525432.
microRNAs (miRNAs) are relevant to metastasis and invasion of non-small cell lung cancer (NSCLC). This study investigated the role of miR-181a-5p in lung cancer. Expression patterns of miR-181a-5p and GTSE1 in the human NSCLC cell line A549 and normal lung epithelial cell line BASE-2B were detected. miR-181a-5p mimic was delivered into A549 cells utilizing Lipofectamine 2000 to overexpress miR-181a-5p, followed by analysis of cell viability, proliferation, apoptosis, invasion, and migration. GTSE1 (G2 and S phase-expressed-1) was predicted as the downstream target gene of miR-181a-5p using bioinformatics analysis software. Targeting relationship between miR-181a-5p and GTSE1 was validated via dual-luciferase assay, RIP assay, and RNA pull-down. Activation of the p53/NF-κB pathway was determined. miR-181a-5p was weakly-expressed in NSCLC cells relative to normal lung epithelial cells. miR-181a-5p overexpression prevented NSCLC cell proliferation, migration, and invasion. Mechanically, miR-181a-5p targeted GTSE1. GTSE1 overexpression partly annulled repression of miR-181a-5p overexpression on NSCLC cell malignant behavior. miR-181a-5p activated the p53 pathway and inhibited the NF-κB pathway by targeting GTSE1. Overall, this study for the first time validated that miR-181a-5p impeded NSCLC cell invasion and migration through activation of the p53 pathway and inhibition of the NF-κB pathway by targeting GTSE1, which may provide a potential novel insight into NSCLC treatment.
微小RNA（miRNA）与非小细胞肺癌（NSCLC）的转移和侵袭相关。本研究调查了miR-181a-5p在肺癌中的作用。检测了miR-181a-5p和GTSE1在人NSCLC细胞系A549和正常肺上皮细胞系BASE-2B中的表达模式。利用Lipofectamine 2000将miR-181a-5p模拟物导入A549细胞以过表达miR-181a-5p，随后分析细胞活力、增殖、凋亡、侵袭和迁移。使用生物信息学分析软件预测GTSE1（G2和S期表达-1）为miR-181a-5p的下游靶基因。通过双荧光素酶测定、RNA免疫沉淀（RIP）测定和RNA下拉实验验证miR-181a-5p与GTSE1之间的靶向关系。测定p53/NF-κB通路的激活情况。相对于正常肺上皮细胞，miR-181a-5p在NSCLC细胞中低表达。miR-181a-5p过表达可抑制NSCLC细胞的增殖、迁移和侵袭。机制上，miR-181a-5p靶向GTSE1。GTSE1过表达部分消除了miR-181a-5p过表达对NSCLC细胞恶性行为的抑制作用。miR-181a-5p通过靶向GTSE1激活p53通路并抑制NF-κB通路。总体而言，本研究首次证实miR-181a-5p通过靶向GTSE1激活p53通路并抑制NF-κB通路来阻碍NSCLC细胞的侵袭和迁移，这可能为NSCLC治疗提供潜在的新见解。
