Du Jun, Zhuo Yingquan, Sun Xu, Nie Meilan, Yang Jiafei, Luo Xi, Gu Huajian
Department of Pediatric Surgery, The Affiliated Hospital of Guizhou Medical University, Guiyang, Guizhou 550004, China.
Department of Pediatric Surgery, The Affiliated Hospital of Guizhou Medical University, No. 16 Beijing Road, Yunyan District, Guiyang, Guizhou 550004, China.
Open Med (Wars). 2023 Jul 14;18(1):20230726. doi: 10.1515/med-2023-0726. eCollection 2023.
Circular RNA has been reported to play a key role in neuroblastoma (NB); however, the role of circ_0000285 in NB remains unclear. The aim of this study was to elucidate the role of circ_0000285 in NB. We studied the expression patterns of miR-582-3p and circ_0000285 in NB tissues and cells using real-time quantitative polymerase chain reaction. The expression of proteins associated with apoptosis (Bax and Bcl-2) and the proteins associated with Wnt/β-catenin (Wnt, p-Gsk-3β, Gsk-3β, β-catenin, and C-myc) were quantified by western blotting. animal models were prepared for the functional verification of circ_0000285 on tumor growth. The potential binding of circ_0000285 to miR-582-3p was ascertained using dual-luciferase reporter and RNA-binding protein immunoprecipitation experiments. Noticeable upregulation of circ_0000285 expression was observed in NB tumor samples and cell lines. and experiments indicated that the absence of circ_0000285 repressed NB cell proliferation and migration, provoked apoptosis, and impaired the activity of Wnt/β-catenin signaling. miR-582-3p is targeted by circ_0000285 and is poorly expressed in NB cells. The additional repression of miR-582-3p in NB cells after circ_0000285 silencing largely recovered circ_0000285 silencing-suppressed NB cell proliferation and migration and enhanced apoptosis. The absence of miR-582-3p restored Wnt/β-catenin signaling activity impaired by the knockdown of circ_0000285. circ_0000285 functions as an miR-582-3p sponge to strengthen Wnt/β-catenin signaling activity, thus exacerbating NB development.
据报道，环状RNA在神经母细胞瘤（NB）中起关键作用；然而，circ_0000285在NB中的作用仍不清楚。本研究的目的是阐明circ_0000285在NB中的作用。我们使用实时定量聚合酶链反应研究了miR-582-3p和circ_0000285在NB组织和细胞中的表达模式。通过蛋白质印迹法定量与凋亡相关的蛋白质（Bax和Bcl-2）以及与Wnt/β-连环蛋白相关的蛋白质（Wnt、p-Gsk-3β、Gsk-3β、β-连环蛋白和C-myc）。制备动物模型以对circ_0000285对肿瘤生长的功能进行验证。使用双荧光素酶报告基因和RNA结合蛋白免疫沉淀实验确定circ_0000285与miR-582-3p的潜在结合。在NB肿瘤样本和细胞系中观察到circ_0000285表达明显上调。 实验表明，circ_0000285缺失会抑制NB细胞增殖和迁移，引发凋亡，并损害Wnt/β-连环蛋白信号传导活性。miR-582-3p是circ_0000285的靶标，在NB细胞中表达较低。circ_0000285沉默后，NB细胞中miR-582-3p的额外抑制在很大程度上恢复了circ_0000285沉默抑制的NB细胞增殖和迁移，并增强了凋亡。miR-582-3p的缺失恢复了因circ_0000285敲低而受损的Wnt/β-连环蛋白信号传导活性。circ_0000285作为miR-582-3p海绵发挥作用，增强Wnt/β-连环蛋白信号传导活性，从而加剧NB的发展。
