Effects of CircRTN4 on the Malignant Biological Behavior of Glioblastoma Cells by Regulating the miR-224-5p/MYT1L Axis
Objective:To investigate the effect of circRNA RTN4(CircRNA)on the malignant biologi-cal 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 Cir-cRTN4 + 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-lucif-erase 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 targe-ting relationships with CircRTN4 and MYT1L.Compared with the blank group and sh NC group,the sh Cir-cRTN4 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 pro-tein expression,decreased apoptosis rate,and cleaved Caspase-3 expression(P<0.05).CircRTN4 expres-sion showed no statistical difference(P>0.05).In vivo experiments demonstrated that interfering with Cir-cRTN4 significantly inhibited tumor mass in nude mice(P<0.05).Conclusion:Silencing CircRTN4 regu-lates the miR-224-5p/MYT1L axis and inhibits the malignant biological behavior of GM cells.
国家科技期刊平台
NSTL
万方数据
维普
