circ_0000885靶向miR-186调控胶质瘤细胞增殖、凋亡和迁移

circ_0000885 regulates the proliferation,apoptosis and migration of glioma cells by targeting miR-186

为探讨环状RNA 0000885(circular RNA 0000885,circ_0000885)对胶质瘤细胞增殖、凋亡和迁移的影响和潜在机制。采用qRT-PCR分析正常脑组织和胶质瘤组织中circ_0000885和miR-186的表达。双荧光素酶报告实验验证circ_0000885和miR-186的靶向关系。将小干扰RNA阴性对照(si-NC)、circ_0000885的小干扰RNA(si-circ_0000885)、miRNA模拟物阴性对照(miR-NC)、miR-186模拟物、miRNA抑制物阴性对照(anti-miR-NC)、miR-186抑制物(anti-miR-186)、si-circ_0000885+anti-miR-186分别转染胶质瘤细胞LN229。通过细胞计数试剂盒(CCK-8)、平板克隆实验、流式细胞术、Transwell实验检测circ_0000885和miR-186表达对LN229细胞活力、集落形成、凋亡和迁移的影响。裸鼠移植瘤实验检测移植瘤体积。结果显示,与正常脑组织比较,胶质瘤组织中circ_0000885的表达水平显著升高(P<0.05),miR-186的表达水平显著降低(P<0.05)。干扰circ_0000885表达或过表达miR-186显著降低LN229细胞活力、集落形成数、迁移数(P<0.05),增加细胞凋亡率和miR-186表达水平(P<0.05),而抑制miR-186表达呈相反作用。抑制miR-186表达可显著减弱干扰circ_0000885表达对LN229细胞活力、集落形成、迁移、凋亡及移植瘤生长的影响(P<0.05)。该研究提示,干扰circ_0000885通过靶向上调miR-186可抑制胶质瘤细胞增殖和迁移,诱导细胞凋亡。

To investigate the effect of circular RNA 0000885(circ_0000885)on the proliferation,apoptosis and migration ofglioma cells,and further to explore its potential mechanism,circ_0000885 and miR-186 expression in normal brain tissues and glioma tissues was analyzed using qRT-PCR.Dual luciferase reporter experiment verified the targeting relationship between circ_0000885 and miR-186.Small interfering RNA negative control(si-NC),circ_0000885 small interfering RNA(si-circ_0000885),miRNA mimic negative control(miR-NC),miR-186 mimic,miRNA inhibitor negative control(anti-miR-NC),miR-186 inhibitor(anti-miR-186),and si-circ_0000885+anti-miR-186 were transfected into glioma cell line LN229.The effects of circ_0000885 and miR-186 expression on cell viability,colony formation,apoptosis and migration of LN229 cells was evaluated by cell counting kit(CCK-8),plate cloning experiment,flow cytometry,and Transwell experiment,respectively.The transplanted tumor volume was detected by the nude mouse xenograft experiment.The results showed that compared to that of the normal brain tissue,circ_0000885 expression in glioma tissue was significantly increased(P<0.05),while miR-186 expression was significantly decreased(P<0.05).Interfering with the expression of circ_0000885 or overexpressing miR-186 significantly decreased the viability,colony formation number,and migration number of LN229 cells(P<0.05),increased cell apoptosis rate and miR-186 expression level(P<0.05),while inhibiting the expression of miR-186 had the opposite effect.Inhibition of miR-186 expression significantly attenuated the effects of JP2interfering circ_0000885 expression on LN229 cell viability,colony formation,migration,apoptosis and xenograft growth.This study suggests that interfering with circ_0000885 inhibits the tumor proliferation and migration,and induces cell apoptosis of glioma cells by targeting and up-regulating miR-186.