circRNA ZNF609靶向miR-512-5p调控胶质瘤细胞恶性生物学行为的分子机制研究  

The Molecular Mechanism of circRNA ZNF609 Targeting miR-512-5p to Regulate the Malignant Biological Behavior of Glioma Cells

作　　者：翟海程[1] 黄琦[1] 郭强[1] ZHAI Hai-cheng;HUANG Qi;GUO Qiang(Department of Neurosurgery,Hanzhong Central Hospital,Hanzhong 723000,China)

机构地区：[1]汉中市中心医院神经外科,汉中723000

出　　处：《中国临床神经科学》2023年第1期1-8,共8页Chinese Journal of Clinical Neurosciences

基　　金：汉中市中心医院院级科研基金(编号:YK1908)。

摘　　要：目的探讨circRNA ZNF609影响胶质瘤细胞恶性生物学行为的可能机制。方法(1)收集35例胶质瘤患者手术切除的肿瘤标本(胶质瘤组)及7例颅脑外伤患者的脑组织标本(对照组),RT-qPCR法检测组织中环状RNA(circRNA)ZNF609与微小RNA(miR)-512-5p表达量;(2)正常胶质细胞(HEB)为对照,RT-qPCR法检测胶质瘤细胞系(LN18、SW1088和Hs683)中circRNA ZNF609与miR-512-5p表达量;(3)胶质瘤LN18细胞,分别转染circRNA ZNF609小干扰RNA(siRNA)、miR-512-5p模拟物、或共转染circRNA ZNF609小干扰siRNA与anti-miRNA-512-5p后,用MTT法、流式细胞术、Transwell法分别检测LN18细胞增殖、凋亡、迁移和侵袭;Western blot法检测细胞中细胞周期蛋白D1(cyclin D1)和cleaved-caspase-3蛋白表达。双荧光素酶报告基因实验验证circRNA ZNF609与miR-512-5p的调控关系。结果与对照组比较,胶质瘤组circRNAZNF609表达升高(t=13.59,P<0.01),miR-512-5p表达降低(t=17.76,P<0.01)。与HEB细胞比较,LN18、SW1088和Hs683细胞中circRNA ZNF609表达量均升高(均P<0.01),miR-512-5p表达量均降低(均P<0.01)。敲减circRNAZNF609或过表达miR-512-5p后,吸光度值、迁移和侵袭数、cyclin D1水平降低,凋亡率和cleaved-caspase-3水平增高(均P<0.01)。结论胶质瘤细胞系中circRNA ZNF609上调表达,敲减circRNA ZNF609表达可能通过负向调控mi R-512-5p表达降低胶质瘤细胞增殖、迁移和侵袭能力,并促进凋亡。Aim To investigate the effect of circRNA ZNF609 on the malignant biological behavior of glioma cells and its possible mechanism.Methods The tumor tissues of 35 patients with glioma and the brain tissues of 7 patients with traumatic brain injury were collected from department of neurosurgery,Hanzhong Central Hospital,and then RT-qPCR method was used to detect the expression of circRNA ZNF609 and miR-512-5p in the tissues.With normal glial cells(HEB)as a control,RT-qPCR was used to detect the expression of circRNA ZNF609 and miR-512-5p in glioma cell lines(LN18,SW1088 and Hs683).Taking glioma LN18 cells as the research object,after circRNA ZNF609 small interfering RNA or miR-512-5p mimic was transfected into LN18 cells,or circRNA ZNF609 small interfering RNA and anti-miR-512-5p were co-transfected into LN18 cells,the cell proliferation,apoptosis,migration and invasion were detected by MTT method,flow cytometry and Transwell,respectively,and Western blotting was used to detect the protein expression of cyclin D1 and cleaved-caspase-3 in cells.The dual luciferase reporter gene experiment verified the regulatory relationship between circRNA ZNF609 and miR-512-5p.Results Compared with the normal brain tissue,the expression of circRNA ZNF609 in glioma tissue was increased(2.65±0.31 vs.1.00±0.17,t=13.58,P<0.01),but the expression of miR-512-5p was decreased(0.45±0.06 vs.1.00±0.13,t=17.76,P<0.01).Compared with HEB,the expression of circRNA ZNF609 in glioma cell lines(LN18,SW1088 and Hs683)increased(P<0.05),and the expression of miR-512-5p decreased(P<0.05).After knocking down circRNA ZNF609 or overexpressing miR-512-5p,the A value,migration number and invasion number,and cyclin D1 level were decreased,while the apoptosis rate and cleaved-caspase-3 level were increased(P<0.05).Conclusion Up-regulation of circRNA ZNF609 in glioma cell lines and knocking down of circRNA ZNF609 expression may reduce the proliferation,migration and invasion ability of glioma cells and promote apoptosis by negatively regulating miR-512-5p e

关 键 词：circRNA ZNF609 miR-512-5p 胶质瘤 细胞增殖 凋亡 迁移 侵袭 

分 类 号：R739.41[医药卫生—肿瘤]