摘要
目的探讨下调hsa_circ_0006988(circ_0006988)对胶质瘤细胞凋亡和迁移的作用。方法实时荧光定量聚合酶链反应(qRT-PCR)检测胶质瘤组织中circ_0006988、miR-3619-5p表达变化。胶质瘤细胞BT325分为Control、si-NC(转染siRNA control)、si-circ(转染circ_0006988 siRNA)、miR-NC(转染mimics control)、miR-3619-5p(转染miR-3619-5p mimics)、si-circ+anti-miR-NC(共转染circ_0006988 siRNA、inhibitor control)、si-circ+anti-miR-3619-5p组(共转染circ_0006988 siRNA、miR-3619-5p inhibitor),qRT-PCR检测miR-3619-5p表达,CCK-8实验检测增殖,流式细胞术检测凋亡,Transwell小室检测迁移,Western Blot检测Bax、B淋巴细胞瘤-2基因(Bcl-2)、基质金属蛋白酶2(MMP-2)蛋白表达。靶基因预测软件预测circ_0006988和miR-3619-5p结合位点,二者关系通过荧光素酶系统确定。结果与瘤旁组织比较,胶质瘤组织中circ_0006988表达水平升高,miR-3619-5p表达水平下降,并且胶质瘤组织中circ_0006988和miR-3619-5p表达水平负相关(r=-0.2897,P<0.001)。与Control、si-NC组比较,si-circ组胶质瘤细胞中miR-3619-5p水平增加,细胞增殖活性、迁移数目以及Bcl-2、MMP-2蛋白表达减少,凋亡率和Bax蛋白表达增多(P<0.05)。与Control、miR-NC组比较,miR-3619-5p组胶质瘤细胞中miR-3619-5p水平增加,细胞增殖活性、迁移数目以及Bcl-2、MMP-2蛋白表达减少,凋亡率和Bax蛋白表达增多(P<0.05)。与si-circ+anti-miR-NC组比较,si-circ+anti-miR-3619-5p组胶质瘤细胞中miR-3619-5p水平降低,细胞增殖活性、迁移数目以及Bcl-2、MMP-2蛋白表达增多,凋亡率和Bax蛋白表达减少(P<0.05)。下调circ_0006988靶向负调控miR-3619-5p表达。结论下调circ_0006988靶向促进miR-3619-5p表达诱导胶质瘤细胞凋亡,并抑制细胞迁移。
Objective To investigate the effect of hsa_circ_0006988(circ_0006988)on apoptosis and migration of glioma cell apoptosis.Methods The expression changes of circ_0006988 and miR-3619-5p in glioma tissues were detected by real-time quantitative polymerase chain reaction(qRT-PCR).Glioma cells BT325 were divided into Control,si-NC(transfected with siRNA control),si-circ(transfected with circ_0006988 siRNA),miR-NC(transfected with mimics control),and miR-3619-5p(transfected with miR-3619-5p mimics),si-circ+anti-miR-NC(co-transfected with circ_0006988 siRNA,inhibitor control),si-circ+anti-miR-3619-5p group(co-transfected with circ_0006988 siRNA,miR-3619-5p inhibitor),qRT-PCR to detect miR-3619-5p expression,CCK-8 assay to detect proliferation,flow cytometry to detect apoptosis,Transwell chamber to detect migration,Western blot to detect Bax,B-cell lymphoma-2(Bcl-2),matrix metalloproteinase 2(MMP-2)protein expression.The target gene prediction software predicted the binding sites of circ_0006988 and miR-3619-5p,and the luciferase reporter system identified the targeting relationship between the two.Results Compared with the paratumor tissue,the expression level of circ_0006988 in glioma tissue was increased,the expression level of miR-3619-5p was decreased,and the expression levels of circ_0006988 and miR-3619-5p in glioma tissue were negatively correlated(r=-0.2897,P<0.001).Compared with the Control and si-NC groups,the level of miR-3619-5p in glioma cells in the si-circ group increased,the cell proliferation activity,the number of migration and the expression of Bcl-2,MMP-2 proteins decreased,and the apoptosis rate and Bax protein increased(P<0.05).Compared with the Control and miR-NC groups,the proliferation activity,migration number,Bcl-2 and MMP-2 protein expressions of glioma cells in the miR-3619-5p group decreased,while the apoptosis rate and Bax protein expression increased(P<0.05).Compared with the si-circ+anti-miR-NC group,the level of miR-3619-5p in the glioma cells of the si-circ+anti-miR-3619-5p group was d
作者
赵水强
孟大伟
陈国强
李亮亮
王志文
胡甜甜
刘东升
牛建星
ZHAO Shuiqiang;MENG Dawei;CHEN Guoqiang(Department of Neurosurgery,Beijing Aviation General Hospital,Beijing 100012,China)
出处
《临床神经外科杂志》
2023年第5期524-530,共7页
Journal of Clinical Neurosurgery
基金
国家自然科学基金青年科学项目(82001377)。
