CK1ε对胶质瘤细胞增殖、迁移和侵袭的影响及其预后相关研究

谢雪露; 武平华

doi:10.16190/j.cnki.45-1211/r.2023.03.002

CK1ε对胶质瘤细胞增殖、迁移和侵袭的影响及其预后相关研究

Effect of CK1ε on proliferation, migration and invasion of glioma cells and its prognosis

摘要

摘要: 目的：探究酪蛋白激酶1ε（CK1ε）通过Wnt/β-catenin 通路对胶质瘤细胞增殖、迁移、侵袭的影响以及CSNK1E（CK1ε 的编码基因）对胶质瘤患者预后的影响。方法：体外培养人胶质瘤U251细胞，转染CK1ε过表达慢病毒载体或阴性对照病毒载体至U251 细胞，分为空白对照组、阴性对照组和实验组。采用细胞计数试剂盒（CCK-8）检测U251 细胞增殖，划痕实验、Transwell 迁移实验检测U251 细胞迁移，Transwell 侵袭实验检测U251 细胞侵袭，蛋白质免疫印迹（western blotting）法检测Wnt/βcatenin 通路相关蛋白β-catenin 蛋白相对表达量。运用Kaplan-Meier 法分析GEPIA 数据库中CSNK1E 表达对脑胶质瘤患者生存期的影响；多因素COX 风险比例回归模型分析中国脑胶质瘤基因组图谱（CGGA）中CSNK1E 表达对脑胶质瘤患者预后的影响。结果：构建了U251-CK1ε过表达体系并且CK1ε成功在U251细胞中过表达，与空白对照组、阴性对照组比较，实验组细胞增殖、迁移数、侵袭数增加（均P＜0.05），β-catenin蛋白相对表达量降低（均P＜0.05）。CSNK1E高表达组患者的总生存期显著比低表达组长（P＜0.05）；高级别胶质瘤、年龄≥50 岁、IDH 突变型是脑胶质瘤患者生存的独立危险因素（均P＜0.05），CSNK1E 表达水平不是脑胶质瘤患者生存的独立影响因素（P＞ 0.05）。结论：CK1ε 能够促进U251 细胞的增殖、迁移和侵袭，可能与抑制Wnt/β-catenin 通路有关；肿瘤恶性程度高、年龄≥50 岁、IDH 突变与胶质瘤患者预后较差有关，而CSNK1E 表达与患者预后无明显关系。

Abstract: Objective: To explore the effect of casein kinase 1ε (CK1ε) on the proliferation, migration and invasion of glioma cells through Wnt/β-catenin pathway and the effect of CSNK1E (CK1ε coding gene) on the prognosis of patients with glioma.Methods: Human glioma U251 cells were cultured in vitro, and transfected with CK1ε overexpressing lentivirus vector or negative virus control, which were divided into blank control group, negative control group and experimental group.Cell counting kit(CCK-8)was used to detect U251 cell proliferation.Scratch test and Transwell test were used to detect U251 cell migration, and Transwell test was used to detect U251 cell invasion as well.Western blotting method was used to detect relative expression of β-catenin protein associated with Wnt/β-catenin pathway.Kaplan-Meier method was used to analyze the effect of CSNK1E expression in GEPIA database on the survival of patients with glioma; multivariate COX risk proportional regression model was used to analyze the effect of CSNK1E expression on the prognosis of patients with glioma in Chinese glioma genome map (CGGA).Results: The overexpression system of U251-CK1ε was established and CK1ε was successfully overexpressed in U251 cells; compared with blank control group and negative control group, the number of cell proliferation, migration and invasion in experimental group increased (all P＜0.05), while the relative expression of β-catenin protein decreased (all P＜0.05).The overall survival of patients with high CSNK1E expression was significantly longer than that with low expression (P＜0.05).High grade gliomas, age higher than 50 years and IDH mutation were independent risk factors for survival of glioma patients (all P＜0.05), while CSNK1E was not an independent factor for survival of glioma patients (P＞ 0.05).Conclusion: CK1ε can promote the proliferation, migration and invasion of U251 cells, which may be related to the inhibition of Wnt/β-catenin pathway; high malignancy, age higher than 50 years, and IDH mutation are associated with poor prognosis in glioma patients, while CSNK1E expression is not significantly correlated with poor prognosis of patients.

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