首页|微小RNA-490-3p靶向高迁移率族蛋白A2/Wnt5a轴抑制脑胶质瘤细胞增殖、迁移机制

微小RNA-490-3p靶向高迁移率族蛋白A2/Wnt5a轴抑制脑胶质瘤细胞增殖、迁移机制

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目的 探讨微小RNA(miRNA,miR)-490-3p抑制脑胶质瘤细胞增殖和迁移的影响及其分子机制.方法 选取2021年7月到2023年7月新乡医学院第一附属医院收治的70例原发性脑胶质瘤标本和癌旁组织作为研究对象,采用荧光定量聚合酶链反应(PCR)分析癌旁组织和胶质瘤组织miR-490-3p的表达水平.复苏脑胶质瘤细胞U251,分为对照组和miR-490-3p组,分别采用脂质体转染对照miRNA和miR-490-3p至U251细胞,转染48 h后,采用细胞计数试剂(CCK-8)和5-乙炔基-2'脱氧尿嘧啶核苷(EdU)染色分析两组细胞增殖能力;体外异体成瘤实验分析两组细胞体内增殖能力;划痕实验和Transwell实验分析两组细胞的迁移和侵袭能力;双荧光素酶报告基因分析miR-490-3p的靶基因,采用蛋白质免疫印迹分析miR-490-3的表达水平.组间计量数据比较采用t检验.结果 脑胶质瘤组织中miR-490-3p水平(0.55±0.11)明显低于癌旁组织(1.18±0.25),差异有统计学意义(t=20.050,P<0.05).对照组细胞CCK-8吸光度值、EdU染色阳性率和肿瘤体积[1.93±0.08、(90.66±4.69)%、(1 085.27±118.17)mm3]明显高于 miR-490-3 p 组[1.47±0.11、(64.12±6.56)%、(685.20±57.28)mm3],差异有统计学意义(t=10.120、10.410、9.634,P<0.05).对照组细胞划痕愈合率和迁移细胞数量[(80.26±5.51)%、(136.60±10.50)个]明显高于miR-490-3p 组[(55.81±6.64)%、(99.50±10.42)个],差异有统计学意义(t=8.958、7.932,P<0.05).高迁移率族蛋白A2(HMGA2)/Wnt5a是miR-490-3p的靶基因.对照组细胞HMGA2和Wnt5a蛋白表达水平(1.12±0.14、1.21±0.12)明显高于 miR-490-3p 组(0.68±0.09、0.75±0.09),差异有统计学意义(t=8.441、10.120,P<0.05).脑胶质瘤组织中HMGA2和Wnt5a蛋白表达水平(1.06±0.14、0.91±0.09)明显高于癌旁组织(1.65±0.18、2.18±0.25),差异有统计学意义(t=21.660、41.030,P<0.05).结论 miR-490-3p在脑胶质瘤组织中呈低表达,通过靶向调节HMGA2/Wnt5a表达水平,调控脑胶质瘤细胞增殖和迁移过程.
Mechanism of microRNA-490-3p targeting high mobility group A2/Wnt5a axis to inhibit prolifera-tion and migration of brain glioma cells
Objective To investigate the effects of microRNA(miRNA,miR)-490-3p on the pro-liferation and migration of brain glioma cells and its molecular mechanism.Methods A total of 70 cases of primary brain glioma specimens and adjacent tissues treated in our hospital from July 2021 to July 2023 were selected as the study objects,and the expression level of miR-490-3p in adjacent tissues and glioma tissues was analyzed by fluorescence quantitative polymerase chain reaction(PCR).Brain glioma cells U251 were resuscitated and divided into control group and miR-490-3p group.Liposomes were transfected into control miRNA and miR-490-3p into U251 cells,respectively.At 48 h after transfection,cell counting kit-8(CCK-8)assay and 5-Ethynyl-2'-deoxyuridine(Edu)staining were used to analyze cell proliferation capacity of the two groups.The in vivo proliferation ability of the two groups of cells was analyzed by alloge-neic tumor formation in vitro.The migration and invasion ability of the two groups of cells was analyzed by scratch test and Transwell test.Bioinformatics and dual luciferase reporter genes were used to analyze the target genes of miR-490-3p,and the expression level of miR-490-3 was analyzed by Western blotting.T test was used to compare measurement data between groups.Results The expression level of miR-490-3p in glioma tissues(0.55±0.11)was significantly lower than that in adjacent tissues(1.18±0.25,t=20.050,P<0.05).CCK-8 absorbance value,EdU staining positive rate and tumor volume[1.93±0.08,(90.66±4.69)%,(1085.27±118.17)mm3]in control group were obviously higher than those in miR490-3p cells[1.47±0.11,(64.12±6.56)%,(685.20±57.28)mm3,t=10.120,10.410,9.634,P<0.05]..The healing rate and number of migrating cells[(80.26±5.51)%,(136.60±10.50)]in control group were obviously higher than those in the miR-490-3p cells[(55.81±6.64)%,(99.50±10.42),t=8.958,7.932,P<0.05].The protein expression levels of HMGA2 and Wnt5a in control group(1.12±0.14,1.21±0.12)were significantly higher than in miR-490-3p group(0.68±0.09,0.75±0.09,t=8.441,10.120,P<0.05).HMGA2 and Wnt5a protein expression levels in brain glioma tissues(1.06±0.14,0.91±0.09)were significantly higher than in paracancer tissue(1.65±0.18,2.18±0.25,t=21.660,41.030,P<0.05).Conclusion MiR-490-3p is low expressed in brain glioma tissues,and it regulates the proliferation and migration of brain glioma cells by targeting and regula-ting the expression level of HMGA2/Wnt5a.

Brain gliomaMicroRNAHigh mobility group A2ProliferationTumor migration

申法政、崔士娟、梁甲宁、王向阳、孟磊、马继伟、赵新利

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新乡医学院第一附属医院神经外科,卫辉 453100

新乡医学院第一附属医院结核内科,卫辉 453100

脑胶质瘤 微小RNA 高迁移率族蛋白A2 增殖 迁移

河南省医学科技攻关计划

LHGJ20210515

2024

中华实验外科杂志
中华医学会

中华实验外科杂志

CSTPCD
影响因子:0.759
ISSN:1001-9030
年,卷(期):2024.41(6)
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