circ_0003645靶向miR-578调控胶质瘤U251细胞增殖、迁移和侵袭的分子机制研究  

The molecular mechanism of circ_0003645 targeting miR-578 to regulate the proliferation,migration and invasion of glioma U251 cells in vitro

作　　者：潘彦波 张春雷[1] 马俊 王昶 PAN Yanbo;ZHANG Chunlei;MA Jun(Department of Neurosurgery,Tieling Central Hospital,Liaoning,Tieling 112000,China)

机构地区：[1]辽宁省铁岭市中心医院神经外科,112000

出　　处：《河北医药》2022年第10期1483-1486,共4页Hebei Medical Journal

摘　　要：目的探讨circ_0003645对胶质瘤U251细胞增殖、迁移和侵袭的影响及分子机制。方法选取35例脑胶质瘤组织及正常脑组织,用实时荧光定量PCR(qRT-PCR)检测circ_0003645和miR-578的表达水平;将胶质瘤细胞株U251随机分为si-circ_0003645组、si-NC组、miR-578组、miR-NC组、si-circ_0003645+anti-miR-578组、si-circ_0003645+anti-miR-NC组;细胞计数试剂盒8(CCK-8)检测细胞活性;克隆形成实验检测细胞克隆形成数;划痕实验检测细胞划痕愈合率;Transwell检测侵袭细胞数;蛋白质印迹(Western blot)法检测蛋白表达;双荧光素酶报告实验检测circ_0003645和miR-578的靶向关系。结果胶质瘤组织中circ_0003645表达水平高于正常脑组织,而miR-578表达水平低于正常脑组织(P<0.05)。与si-NC组比较,si-circ_0003645组circ_0003645表达水平降低,细胞活性降低,细胞克隆形成数减少,划痕愈合率降低,侵袭细胞数减少,E-cadherin表达水平升高,N-cadherin表达水平降低(P<0.05)。过表达miR-578后,miR-578表达水平升高,U251细胞的活性降低,细胞克隆形成数减少,划痕愈合率降低,侵袭细胞数减少,E-cadherin表达水平升高,N-cadherin表达水平降低(P<0.05)。Circular RNA Interactome预测显示circ_0003645与miR-578含有互补的核苷酸序列。WT-circ_0003645与miR-578共转染的U251细胞荧光素酶活性降低(P<0.05);过表达circ_0003645后,miR-578表达水平降低;抑制circ_0003645表达后,miR-578表达水平升高(P<0.05)。与si-circ_0003645+anti-miR-NC组比较,si-circ_0003645+anti-miR-578组miR-578表达水平降低,细胞活性升高,细胞克隆形成数增加,划痕愈合率升高,侵袭细胞数增加,E-cadherin表达水平降低,N-cadherin表达水平升高(P<0.05)。结论干扰circ_0003645表达通过靶向上调miR-578抑制胶质瘤U251细胞增殖、迁移和侵袭。Objective To investigate the effects of circ_0003645 on the proliferation,migration and invasion of glioma U251 cells in vitro,and to explore its molecular mechanism.Methods A total of 35 cases of glioma tissues and normal brain tissues were selected,and the expression levels of circ_0003645 and miR-578 were detected by Real-time fluorescent quantitative PCR(qRT-PCR).The glioma cell line U251 was randomly divided into si-circ_0003645 group,si-NC group,miR-578 group,miR-NC group,si-circ_0003645+anti-miR-578 group,si-circ_0003645+anti-miR-NC group.The cell counting kit 8(CCK-8)was used to detecte cell viability;clone formation experiment detected the number of cell clone formation;the scratch experiment detected the cell scratch healing rate;Transwell detected the number of invaded cells;Western Blot detected protein expression;the dual luciferase reporter experiment detected the targeting correlation of circ_0003645 and miR-578.Results The expression levels of circ_0003645 in glioma tissues were significantly higher than those in normal tissues,however,the expression levels of miR-578 were significantly lower than those in normal tissues(P<0.05).Compared with those in si-NC group,the expression levels of circ_0003645,cell activity,scratch healing rate,the number of cell clone formation and the number of invasive cells as well as the expression levels of N-cadherin in si-circ_0003645 group were significantly decreased,however,the expression levels of E-cadherin were significantly increased(P<0.05).The circ_0003645 targeted and regulated the expression of miR-578,and down-regulating the expression of miR-578 reversed the effects of interfering with the expression of circ_0003645 on the proliferation,migration and invasion of glioma U251 cells.Conclusion Interfering the expression of circ_0003645 can inhibit the proliferation,migration and invasion of glioma U251 cells by targetedly up-regulating the expression of miR-578.

关 键 词：circ_0003645 miR-578 胶质瘤 增殖 迁移 侵袭 

分 类 号：R739.41[医药卫生—肿瘤]