敲减SNORA31抑制胶质瘤干细胞血管生成拟态的作用机制

Mechanisms of SNORA31-knockdown inhibiting vasculogenic mimicry of glioma stem cells

目的探讨SNORA31在胶质瘤干细胞(GSCs)中的表达以及抑制GSCs血管生成拟态的作用机制。方法对中国脑胶质瘤基因组图谱(CGGA)数据库中的患者预后数据与SNORA31表达量进行生存分析。采用qRT-PCR检测GSCs中SNORA31表达。敲减SNORA31后Western blotting检测促红细胞生成素肝细胞受体A2(EphA2)表达,细胞外调节蛋白激酶1/2(ERK1/2)和磷酸肌醇3激酶(PI3K)表达与磷酸化水平;CCK8和Transwell检测GSCs增殖、迁移和侵袭的变化;基质胶三维管形成实验检测血管生成拟态能力的变化。结果与亲本细胞比较,SNORA31在GSCs中的表达显著增高(P<0.05)。与相应的阴性对照组比较,敲减SNORA31显著降低EphA2的表达水平和p-ERK1/2/ERK1/2、p-PI3K/PI3K比值(均P<0.05),进而抑制GSCs的增殖、迁移、侵袭和血管生成拟态的形成。结论敲减SNORA31降低了EphA2表达水平、抑制了ERK1/2和PI3K磷酸化水平,进而抑制GSCs的增殖、迁移、侵袭和血管生成拟态的形成。

Objective The present study aimed to investigate the expression of SNORA31 in glioma stem cells(GSCs)and the influence of the regulation of erythropoietin-producing hepatocellular receptor A2(EphA2)on the proliferation,migration,invasion,and vasculogenic mimicry of GSCs.Methods Survival analysis was performed on SNORA31 expression and patient prognosis data obtained from the(Chinese gliomagenome atlas,CGGA)database.The expression of SNORA31 in GSCs was detected using qRT-PCR.After SNORA31 knockdown,the expression of EphA2 and phosphorylation of ERK1/2 and PI3K were detected by Western blotting.CCK8 and Transwell assays were used to examine alterations in the proliferation,migration,and invasion of GSCs.A matrigel three-dimensional tube formation test was used to detect changes in vasculogenic mimicry.Results Compared with parental cells,the expression of SNORA31 was significantly increased in GSCs(P<0.05).Knockdown of SNORA31 significantly reduced the expression of EphA2,p-ERK1/2/ERK1/2,and p-PI3K/PI3K and inhibited the proliferation,migration,invasion and vasculogenic mimicry of GSCs.Conclusion Knockdown of SNORA31 reduces the phosphorylation of ERK1/2 and PI3K by downregulating EphA2 expression,thereby inhibiting the proliferation,migration,invasion,and vasculogenic mimicry of GSCs.