The Third Department of Surgery, The First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China.
Bioengineered. 2021 Dec;12(2):12880-12890. doi: 10.1080/21655979.2021.2005986.
We investigated the effect of the long noncoding RNA (lncRNA) maternally expressed gene 3 (MEG3) on hepatocellular carcinoma (HCC) tumorigenesis and progression by targeting miR-5195-3p and transcription factor forkhead box O1 (FOXO1) to identify a novel target for HCC treatment. HCC clinical samples were collected, and cell counting kit-8 (CCK-8), and transwell migration and invasion assays were performed. Furthermore, interaction was detected via double luciferase reporter and RNA pull-down assays. MEG3, miR-5195-3p, and FOXO1 expression was determined by quantitative real-time polymerase chain reaction (RT-qPCR) and Western blotting. Xenograft tumor models were established to investigate the effect of MEG3 . Compared with normal tissues, MEG3 expression was significantly downregulated in HCC tissues. MEG3 overexpression inhibited the viability and migration of HCC cells. Double luciferase reporter and RNA pull-down assays confirmed the binding between MEG3 and miR-5195-3p as well as between miR-5195-3p and FOXO1. RT-qPCR and Western blotting results showed that MEG3 inhibited the expression of miR-5195-3p and promoted that of FOXO1. Additionally, MEG3 overexpression inhibited HCC tumorigenesis and progression in xenograft tumor models while depletion of MEG3 exerted the opposite way. Therefore, the lncRNA MEG3 inhibits HCC tumorigenesis and progression through the miR-5195-3p/FOXO1 signaling axis.
我们通过靶向 microRNA-5195-3p 和转录因子叉头框 O1（FOXO1）来研究长链非编码 RNA（lncRNA）母系表达基因 3（MEG3）对肝细胞癌（HCC）发生和发展的影响，以确定 HCC 治疗的新靶点。收集 HCC 临床样本，进行细胞计数试剂盒-8（CCK-8）、Transwell 迁移和侵袭实验。此外，通过双荧光素酶报告和 RNA 下拉实验检测相互作用。通过定量实时聚合酶链反应（RT-qPCR）和 Western blot 检测 MEG3、miR-5195-3p 和 FOXO1 的表达。建立异种移植肿瘤模型以研究 MEG3 的作用。与正常组织相比，MEG3 在 HCC 组织中的表达明显下调。MEG3 过表达抑制 HCC 细胞的活力和迁移。双荧光素酶报告和 RNA 下拉实验证实了 MEG3 与 miR-5195-3p 以及 miR-5195-3p 与 FOXO1 之间的结合。RT-qPCR 和 Western blot 结果表明，MEG3 抑制了 miR-5195-3p 的表达并促进了 FOXO1 的表达。此外，MEG3 过表达抑制了异种移植肿瘤模型中的 HCC 肿瘤发生和进展，而 MEG3 的耗竭则产生了相反的效果。因此，lncRNA MEG3 通过 miR-5195-3p/FOXO1 信号通路抑制 HCC 肿瘤发生和发展。
