circPRMT5靶向miR-376a-3p调控胶质瘤U251细胞增殖和凋亡的作用机制  

circPRMT5 Targets miR-376a-3p to Regulate Proliferation and Apoptosis of Glioma U251 Cells

作　　者：黄超 方兴刚 陈璐 HUANG Chao;FANG Xinggang;CHEN Lu(Taihe Hospital,Affiliated Hospital of Hubei University of Medicine,Shiyan 442000,Hubei,China)

机构地区：[1]十堰市太和医院/湖北医药学院附属医院,湖北十堰442000

出　　处：《中西医结合心脑血管病杂志》2025年第4期547-554,共8页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：2022年度湖北省教育厅科学研究计划指导性项目(No.B2022131)。

摘　　要：目的:探讨circPRMT5对胶质瘤U251细胞增殖和凋亡的影响及分子机制。方法:选取2020年5月—2022年5月我院脑胶质瘤组织和颅脑外伤病人脑组织标本各37例;胶质瘤细胞U251分为si-NC组、si-circPRMT5组、pcDNA组、pcDNA-circPRMT5组、miR-NC组、miR-376a-3p组、si-circPRMT5+anti-miR-NC组及si-circPRMT5+anti-miR-376a-3p组。采用实时荧光定量PCR(qRT-PCR)检测circPRMT5和miR-376a-3p表达水平;四甲基偶氮唑盐比色法(MTT)检测细胞增殖活性;流式细胞仪检测细胞凋亡;蛋白质免疫印迹法(Western Blot)检测细胞增殖及凋亡相关蛋白表达。双荧光素酶报告实验检测circPRMT5和miR-376a-3p的靶向调控关系。结果:与颅脑外伤病人脑组织比较,胶质瘤组织中circPRMT5表达水平(2.55±0.18与1.00±0.07)升高,miR-376a-3p表达水平[(0.42±0.05与1.00±0.06)]降低(P<0.05)。抑制circPRMT5表达后,细胞活性(0.32±0.03与0.74±0.06)降低,细胞凋亡率[(22.37±1.83)%与(7.47±0.62)%]升高,周期蛋白D1(CyclinD1)及B细胞淋巴瘤/白血病2(Bcl-2)蛋白相对表达量降低,细胞周期蛋白依赖性激酶抑制剂1A(p21)及Bcl-2相关x蛋白(Bax)相对表达量升高(P<0.05)。过表达miR-376a-3p后,细胞活性(0.41±0.04与0.76±0.07)降低,细胞凋亡率[(19.51±1.40)%与(7.37±0.53)%]升高,CyclinD1及Bcl-2蛋白相对表达量降低,p21及Bax蛋白相对表达量升高(P<0.05)。miR-376a-3p组转染WT-circPRMT5的细胞荧光素酶活性显著低于miR-NC组(0.59±0.04与1.03±0.07,P<0.05),而转染MUT-circPRMT5的细胞荧光素酶活性无显著变化(1.00±0.05与1.02±0.08,P>0.05)。结论:抑制circPRMT5表达可能通过靶向上调miR-376a-3p抑制胶质瘤U251细胞增殖,促进细胞凋亡。Objective:To explore the effect and molecular mechanism of circPRMT5 on the proliferation and apoptosis of glioma U251 cells.Methods:A total of 37 specimens of brain glioma tissues and 37 specimens of brain tissues from patients with craniocerebral trauma were selected from May 2020 to May 2022.U251 glioma cells were divided into si-NC group,si-circPRMT5 group,pcDNA group,pcDNA-circPRMT5 group,miR-NC group,miR-376a-3p group,si-circPRMT5+anti-miR-NC group,and si-circPRMT5+anti-miR-376a-3p group.Real-time fluorescence quantitative PCR(RT-qPCR)was used to detect the expressions of circPRMT5 and miR-376a-3p.Methyl thiazolyl tetrazolium(MTT)was used to detect cell activity.Flow cytometry was used to detect apoptosis.The expression of proteins related to cell proliferation and apoptosis was detected by Western Blot.The dual luciferase reporter assay was used to detect the relationship between circPRMT5 and miR-376a-3p.Results:Compared with the brain tissue of patients with traumatic brain injury,the expression level of circPRMT5 in glioma tissue was increased(2.55±0.18 vs 1.00±0.07,P<0.05),and the expression level of miR-376a-3p was decreased(0.42±0.05 vs 1.00±0.06,P<0.05).After inhibiting the expression of circPRMT5,cell viability(0.32±0.03 vs 0.74±0.06)was decreased,cell apoptosis rate[(22.37±1.83)%vs(7.47±0.62)%]was increased,the expression levels of CyclinD1 and B-cell lymphoma/leukemia-2(Bcl-2)were decreased,and the expression levels of cyclin-dependent kinase inhibitor 1A(p21)and Bcl-2 related X(Bax)protein were increased(P<0.05).After overexpression of miR-376a-3p,cell viability(0.41±0.04 vs 0.76±0.07)was decreased,and cell apoptosis rate[(19.51±1.40)%vs(7.37±0.53)%]was increased,the expression levels of CyclinD1 and Bcl-2 were decreased,and the expression levels of p21 and Bax were increased(P<0.05).Compared with the miR-NC group,the luciferase activity of cells transfected with WT-circPRMT5 in the miR-376a-3p group was decreased(0.59±0.04 vs 1.03±0.07,P<0.05),while the luciferase activity of cell

关 键 词：胶质瘤 增殖 凋亡 circPRMT5 miR-376a-3p 实验研究 

分 类 号：R73[医药卫生—肿瘤]