沉默circ_0008450通过调控miR-338-3p/FOXP2信号通路抑制胶质瘤细胞的发生发展

Silencing circ_0008450 Inhibits the Occurrence and Development of Glioma Cell U251 by Regulating miR-338-3p/FOXP2

目的 探讨沉默circ_0008450对胶质瘤细胞(U251)发生发展的影响及分子机制。方法 选取31例胶质瘤组织并依据胶质瘤病理分级分为低分级组(Ⅰ-Ⅱ期,15例)和高分级组(Ⅲ-Ⅳ期,16例),另选择25例内减压术获取的正常脑组织标本为正常对照组。运用实时荧光定量PCR(RT-qPCR)和Western blot法分别检测circ_0008450、miR-338-3p和FOXP2蛋白的表达水平。将U251细胞分为对照组、si-NC组、si-circ_0008450组、miR-NC组、miR-338-3p mimic组、si-circ_0008450+miR-338-3p inhibitor组。使用MTT法、克隆形成实验、流式细胞术和划痕实验分别检测不同转染的U251细胞的增殖抑制率、集落形成数、细胞凋亡及划痕愈合率;双荧光素酶报告实验检测circ_0008450、miR-338-3p及FOXP2之间的靶向关系。结果 与正常对照组比较,低分级组和高分级组的胶质瘤组织中circ_0008450和FOXP2表达水平升高,miR-338-3p表达水平降低(均P<0.05);circ_0008450和FOXP2与miR-338-3p表达水平均呈负相关(均P<0.05)。沉默circ_0008450或过表达miR-338-3p后FOXP2表达水平降低、U251细胞增殖抑制率升高、细胞凋亡率升高、划痕愈合率降低、集落形成数减少(均P<0.05)。circ_0008450靶向调控miR-338-3p;miR-338-3p靶向调控FOXP2。抑制miR-338-3p可消除circ_0008450沉默对U251增殖、迁移、凋亡的影响。结论 沉默circ_0008450通过调控miR-338-3p/FOXP2轴抑制U251细胞增殖和迁移,促进细胞凋亡。

Aim To explore the influence and molecular mechanism of circ_0008450 on the occurrence and development of glioma cell U251.Methods Thirty-one cases of glioma tissues and 25 cases of normal brain tissue specimens were selected,and 31 glioma tissues were divided into a low grade group(stageⅠ-Ⅱ,n=15)and a high grade group(stageⅢ-Ⅳ,n=16).The expression levels of circ_0008450,miR-338-3p and FOXP2 protein were detected by real-time fluorescent quantitative PCR(RT-qPCR)and Western blotting,respectively.The glioma cells U251 were divided into a Con group,a si-NC group,a si-circ_0008450 group,a miR-NC group,a miR-338-3p mimic group,a si-circ_0008450+miR-338-3p inhibitor group.The proliferation inhibition rate,colony formation number,apoptosis and scratch healing rate of U251 cells transfected with different transfections were assessed by tetramethylazolium salt colorimetry(MTT),colony formation assay,flow cytometry and scratch assay,respectively.The targeting relationship among circ_0008450,miR-338-3p and FOXP2 were detected by dual luciferase report test.Results Compared with the normal brain tissue,the expression levels of circ_0008450 and FOXP2 in glioma tissues of the low grade group and high grade group were increased,and the expression level of miR-338-3p was decreased,and the difference between the two groups was significant.The expression levels of circ_0008450,FOXP2 and miR-338-3p were negatively correlated(P<0.05).After silencing circ_0008450 or overexpressing of miR-338-3p,FOXP2 expression level was decreased,and U251 cell proliferation inhibition rate,apoptosis rate were increased,scratch healing rate and the colony formation number were decreased(P<0.05).Circ_0008450 targeted and regulated miR-338-3p;miR-338-3p targeted and regulated FOXP2.Inhibition of miR-338-3p can eliminate the effect of circ_0008450 silencing on the proliferation,migration and apoptosis of U251.Conclusion Silencing circ_0008450 inhibits the proliferation and migration of glioma cells U251 and promotes cell apoptosis by regulating the miR-338-3p/FOXP2 axis.