CircRTN4调节miR-224-5p/MYT1L轴对胶质母细胞瘤细胞恶性生物学行为的影响  

Effects of CircRTN4 on the Malignant Biological Behavior of Glioblastoma Cells by Regulating the miR-224-5p/MYT1L Axis

作　　者：刘应刚 李维民[1] 龙勇[1] 向城卫 张施远[1] 陈星宇 LIU Yinggang;LI Weimin;LONG Yong(Suining Central Hospital,Sichuan Suining 629018,China)

机构地区：[1]四川省遂宁市中心医院,四川遂宁629018

出　　处：《河北医学》2024年第1期15-22,共8页Hebei Medicine

基　　金：四川省医学(青年创新)科研课题立项项目,(编号:Q21028)。

摘　　要：目的:探讨环状RNA(CircRNA)RTN4调节微小RNA(miR)-224-5p/髓磷脂转录因子1样(MYT1L)轴对胶质母细胞瘤(GM)细胞恶性生物学行为的影响。方法:以对数生长期的U251细胞进行设置分组:空白组、阴性对照(sh NC)组、沉默circRTN4 shRNA(sh circRTN4)组、sh circRTN4+抑制剂对照(inhibitor NC)组、sh circRTN4+miR-224-5p抑制剂(miR-224-5p inhibitor)组。Transwell实验、流式细胞术、CCK-8、qRT-PCR、Western blot以及双荧光素酶分别检测细胞迁移与侵袭、凋亡、增殖、CircRTN4、miR-224-5p、MYT1L mRNA表达、增殖蛋白(ki-67)、凋亡蛋白(cleaved Caspase-3)、MYT1L蛋白表达以及miR-224-5p分别与CircRTN4、MYT1L靶向关系验证;qRT-PCR检测GM细胞系U118、U87、U251和LN229细胞及正常人星形胶质细胞系NHA细胞中CircRTN4、miR-224-5p、MYT1L mRNA表达水平。结果:U118、U87、U251和LN229细胞中CircRTN4、MYT1L mRNA表达较NHA细胞显著增加,miR-224-5p表达显著下降(P<0.05);miR-224-5p分别与CircRTN4、MYT1L的有靶向关系。与空白组、sh NC组相比,sh circRTN4组迁移、侵袭数、24h、48h A450值、ki-67、CircRTN4、MYT1L mRNA及蛋白表达显著降低,凋亡率、cleaved Caspase-3表达显著增加(P<0.05);与sh circRTN4+inhibitor NC组相比,sh circRTN4+miR-224-5p inhibitor组迁移、侵袭数、24h、48h A450值、ki-67、MYT1L mRNA及蛋白表达显著增加,凋亡率、cleaved Caspase-3表达显著下降(P<0.05),CircRTN4表达无统计学差异(P>0.05),体内实验证明干扰CircRTN4可显著抑制移植瘤裸鼠肿瘤质量(P<0.05)。结论:沉默CircRTN4调节miR-224-5p/MYT1L轴抑制GM细胞恶性生物学行为。Objective:To investigate the effect of circRNA RTN4(CircRNA)on the malignant biological behavior of glioblastoma(GM)cells by regulating miR-224-5p/myelin transcription factor-1-like(MYT1L)axis.Methods:U251 cells in logarithmic growth phase were divided into the following groups:blank group,negative control(sh NC)group,silenced CircRTN4 shRNA(sh CircRTN4)group,sh CircRTN4+inhibitor control(inhibitor NC)group,and sh CircRTN4+miR-224-5p inhibitor(miR-224-5p inhibitor)group.Transwell experiment,flow cytometry,CCK-8,qRT-PCR,Western blot,and dual-luciferase assay were used to detect cell migration and invasion,apoptosis,proliferation,CircRTN4,miR-224-5p,MYT1L mRNA expression,proliferation protein(ki-67),apoptosis protein(cleaved Caspase-3),and MYT1L protein expression.The targeting relationships between miR-224-5p and CircRTN4,MYT1L were verified.qRT-PCR was used to detect the mRNA expression levels of CircRTN4,miR-224-5p,and MYT1L in GBM cell lines(U118,U87,U251,LN229)and normal human astrocytes(NHA)cells.Results:In U118,U87,U251,and LN229 cells,CircRTN4 and MYT1L mRNA expression increased significantly,while miR-224-5p expression decreased significantly compared to NHA cells(P<0.05).miR-224-5p had targeting relationships with CircRTN4 and MYT1L.Compared with the blank group and sh NC group,the sh CircRTN4 group showed significantly decreased migration,invasion,24h,48h A450 values,ki-67,CircRTN4,MYT1L mRNA,and protein expression,increased apoptosis rate,and cleaved Caspase-3 expression(P<0.05).Compared with the sh CircRTN4+inhibitor NC group,the sh CircRTN4+miR-224-5p inhibitor group showed significantly increased migration,invasion,24h,48h A450 values,ki-67,MYT1L mRNA,and protein expression,decreased apoptosis rate,and cleaved Caspase-3 expression(P<0.05).CircRTN4 expression showed no statistical difference(P>0.05).In vivo experiments demonstrated that interfering with CircRTN4 significantly inhibited tumor mass in nude mice(P<0.05).Conclusion:Silencing CircRTN4 regulates the miR-224-5p/MYT1L axis and inhibits the ma

关 键 词：胶质母细胞瘤 环状RTN4 miR-224-5p/MYT1L轴 恶性生物学行为 

分 类 号：R739.41[医药卫生—肿瘤]