miRNA-128-3p通过下调KLHDC8A的表达抑制胶质瘤细胞的恶性生物学行为  

miRNA-128-3p inhibits malignant behavior of glioma cells by downregulating KLHDC8A expression

作　　者：于正涛[1] 李佳梦 蒋俊文 李由 林珑 夏鹰[1] 王磊 YU Zhengtao;LI Jiameng;JIANG Junwen;LI You;LIN Long;XIA Ying;WANG Lei(Department of Neurosurgery,Affiliated Haikou Hospital of Xiangya School of Central South University,Haikou 570208,China;Department of Neurosurgery,Hunan Cancer Hospital and Affiliated Cancer Hospital of Xiangya School of Medicine,Central South University,Changsha 410006,China)

机构地区：[1]中南大学湘雅医学院附属海口医院神经外科,海南海口570208 [2]中南大学湘雅医学院附属肿瘤医院神经外科,湖南长沙410006

出　　处：《南方医科大学学报》2023年第9期1447-1459,共13页Journal of Southern Medical University

基　　金：海南省自然科学基金(820MS163);湖南省卫生健康委员会科研项目(20200709);海南省脑血管病临床医学研究中心资助项目(LCYX202206)。

摘　　要：目的探讨miRNA-128-3p在胶质瘤细胞中的作用,并确定其是否能够调控KLHDC8A介导恶性生物学行为,为寻找胶质瘤患者潜在的分子生物标志物和治疗靶点提供理论支持。方法采用双荧光素酶报告基因、qRT-PCR和Western blotting实验验证miRNA-128-3p与KLHDC8A的调控关系。通过Edu实验、流式细胞术、Transwell和划痕实验验证miRNA-128-3p和KLHDC8A对胶质瘤细胞恶性行为的影响。利用挽救实验验证miRNA-128-3p靶向KLHDC8A调控胶质瘤细胞的增殖、凋亡、侵袭和迁移。结果KLHDC8A在高级别胶质瘤组织中的表达水平显著升高,与恶性胶质瘤患者的生存状况密切相关。功能验证实验表明,高表达KLHDC8A可促进胶质瘤细胞的增殖、迁移和侵袭,而miRNA-128-3p高表达抑制胶质瘤细胞的增殖、迁移等恶性生物学行为。miRNA-128-3p靶向调节胶质瘤细胞中KLHDC8A的表达,miRNA-128-3p高表达通过靶向KLHDC8A抑制胶质瘤细胞的恶性生物学行为。结论miRNA-128-3p负向调控其下游靶基因KLHDC8A的表达,抑制胶质瘤细胞恶性生物学行为,因此miRNA-128-3p/KLHDC8A是判断胶质瘤预后及治疗的分子靶点。Objective To determine whether miRNA-128-3p regulates malignant biological behavior of glioma cells by targeting KLHDC8A.Methods Dual-luciferase reporter assays,qRT-PCR andWestern blotting were used to verify the targeting of miRNA-128-3p to KLHDC8A.Edu assay,flow cytometry,Transwell assay and would healing assay were used to determine the effects of changes in miRNA-128-3p and KLHDC8Aexpression levels on malignant behavior of glioma cells.Rescue experiment was carried out to verify that miRNA-128-3p regulated glioma cell proliferation,apoptosis,invasion and migration by targeting KLHDC8A.Results The expression level of KLHDC8Awas significantly increased in high-grade glioma tissue andwas closely related to a poor survival outcome of the patients.Overexpression of KLHDC8A promoted glioma cell proliferation,migration and invasion,and miRNA-128-3p overexpression inhibited proliferative and metastatic capacities of glioma cells.Mechanistically,KLHDC8A expression was directly modulated by miRNA-128-3p,which,by targeting KLHDC8A,inhibited malignant behavior of glioma cells.Conclusion Upregulation of miRNA-128-3p inhibits uncontrolled growth of glioma cells by negatively regulating KLHDC8A expression and its downstream effectors,suggesting that the miRNA-128-3p-KLHDC8A axis may serve as a potential prognostic indicator and a therapeutic target for developing new strategies for glioma treatment.

关 键 词：胶质瘤 miRNA-128-3p KLHDC8A 生物标志物 迁移 增殖 

分 类 号：R739.4[医药卫生—肿瘤]