Drug Clinical Trial Institution, The First Hospital of Shanxi Medical University, Taiyuan, 030001, Shanxi Province, China.
Clinical Laboratory, Taiyuan People's Hospital, Taiyuan, 030001, Shanxi Province, China.
Cell Mol Biol (Noisy-le-grand). 2023 Jun 30;69(6):132-140. doi: 10.14715/cmb/2023.69.6.20.
Circular RNA (circRNA) is a kind of RNA generated by a covalently closed loop and possesses sophisticated capacities of gene regulation in tumorigenesis and development. However, the role of circZSWIM4 on lung adenocarcinoma (LUAD) remains largely unclear. In the present study, we used reverse transcription-qPCR (RT-qPCR) to examine whether circZSWIM4 was significantly overexpressed in LUAD cells. The impacts of circZSWIM4 on the properties of proliferation, apoptosis and migration were assessed by loss-of and gain-of-function assays, such as CCK-8 experiments, flow cytometry analysis and wound healing experiments. Moreover, TOP/FOP flash experiments and FISH experiments were carried out to prove that circZSWIM4 stimulated the Wnt/β-catenin pathway. Downstream targets of circZSWIM4 were forecasted by bioinformatics tools and validated by RNA immunoprecipitation (RIP), RNA pulls down as well as luciferase reporter experiments. Forkhead box M1 (FOXM1) was confirmed to be the corporate targets of miR-370-3p and miR-873-5p. Through co-IP assay, we verified the combination between FOXM1 and β-catenin. Totally, circZSWIM4 activated the Wnt/β-catenin pathway by targeting miR-370-3p and miR-873-5p to regulate FOXM1 and β-catenin and facilitated the progression of LUAD.
环状 RNA(circRNA)是一种通过共价闭环产生的 RNA,具有在肿瘤发生和发展中进行基因调控的复杂能力。然而,circZSWIM4 在肺腺癌(LUAD)中的作用在很大程度上仍不清楚。在本研究中,我们使用逆转录-qPCR(RT-qPCR)来检测 circZSWIM4 是否在 LUAD 细胞中显著过表达。通过缺失和获得功能测定(如 CCK-8 实验、流式细胞术分析和划痕愈合实验)评估 circZSWIM4 对增殖、凋亡和迁移特性的影响。此外,进行 TOP/FOP 闪烁实验和 FISH 实验以证明 circZSWIM4 刺激 Wnt/β-catenin 通路。通过生物信息学工具预测 circZSWIM4 的下游靶标,并通过 RNA 免疫沉淀(RIP)、RNA 下拉和荧光素酶报告基因实验进行验证。叉头框 M1(FOXM1)被证实是 miR-370-3p 和 miR-873-5p 的共同靶标。通过 co-IP 实验,我们验证了 FOXM1 和 β-catenin 之间的结合。总的来说,circZSWIM4 通过靶向 miR-370-3p 和 miR-873-5p 来调节 FOXM1 和 β-catenin,从而激活 Wnt/β-catenin 通路,促进 LUAD 的进展。