极光激酶A抑制剂对胶质母细胞瘤细胞增殖和B7-H3蛋白表达影响的实验研究  

Experimental study on the effects of AURKA inhibitor on glioblastoma cell proliferation and B7-H3 proteinexpression

作　　者：刘金秋 蒲焯楠 邓宇轩 吕一帆 张绍东[1] 郝淑煜[2] 季楠[2] 万虹[1] 冯洁[1] Liu Jinqiu;Pu Zhuonan;Deng Yuxuan;Lyu Yifan;Zhang Shaodong;Hao Shuyu;Ji Nan;Wan Hong;Feng Jie(Beijing Neurosurgical Instute,Capital Medical University,Beijing 100070,China;Neurosurgery Center Beijing Tiantan Hospital,Capital Medical University,Beijing 100070,China)

机构地区：[1]首都医科大学,北京市神经外科研究所,北京100070 [2]首都医科大学附属北京天坛医院神经外科学中心,北京100070

出　　处：《中华神经外科杂志》2024年第4期406-413,共8页Chinese Journal of Neurosurgery

基　　金：国家自然科学基金(81872052)。

摘　　要：目的探讨极光激酶A(AURKA)抑制剂对胶质母细胞瘤(GBM)的细胞增殖和免疫检查点B7-H3表达的影响。方法基于中国胶质瘤基因组图谱(CGGA)计划数据库(共325例患者),分析AURKA mRNA的表达水平与肿瘤的病理学类型、世界卫生组织(WHO)级别、原/复发状态、患者的生存期、O°-甲基鸟嘌呤-DNA-甲基转移酶(MGMT)甲基化、异柠檬酸脱氢酶(IDH)突变及染色体1p19q共缺失的相关性。比较AURKA mRNA在不同病理学类型、不同WHO级别,以及原发与复发性脑胶质瘤中表达的差异。绘制Kaplan-Meier生存曲线以比较不同AURKA mRNA表达水平的脑胶质瘤患者生存期差异。通过单因素和多因素Cox回归模型分析探讨AURKAmRNA表达水平、IDH突变、染色体1p19q共缺失、发病年龄、放疗、化疗、WHO级别及原/复发状态对脑胶质瘤患者生存期的影响。绘制受试者工作特征(ROC)曲线并计算曲线下面积(AUC),以确定AURKA mRNA表达水平对脑胶质瘤患者生存期的预测价值。收集首都医科大学附属北京天坛医院神经外科学中心接受手术治疗患者的脑胶质瘤组织样本,其中低级别胶质瘤(LGG)4例,CBM4例;通过蛋白质免疫印迹(WB)法检测AURKA在肿瘤组织样本中的表达情况。采用AURKA抑制剂alisertib处理人CBM细胞株U87-MG,将其分为对照组(以DMSO处理)和alisertib处理组(以5μmol/Lalisertib处理)。采用CCK-8法和结晶紫染色方法分别检测alisertib对细胞活性和克隆形成的影响。采用WB法检测alisertib对AURKA、磷酸化AURKA和B7-H3蛋白表达的影响。应用流式细胞术检测alisertib对GBM细胞膜蛋白B7-H3表达的影响。结果CGGA计划数据库分析结果表明,AURKA mRNA在CBM中的表达水平高于星形胶质细胞瘤和少突胶质细胞瘤(均P<0.001);在WHO2、3及4级胶质瘤组间,AURKA mRNA的表达水平随WHO级别的升高而增加(P<0.001);AURKA mRNA在复发性胶质瘤中的表达水平高于原发性胶质瘤(t=4.50,P<0.001)。KObjective To investigate the effects of aurora kinase A(AURKA)inhibitor on the proliferation of glioblastoma(GBM)cells and expression of immune checkpoint B7-H3.Methods The Chinese Glioma Genome Atlas(CGGA)dataset(325 cases)was used to analyze the correlation between expression levels of AURKA mRNA and clinical data including histopathology,World Health Organization(WHO)grade,primary/recurrent status,overall survival(OS),O°-methylguanine-DNA-methyltransferase(MGMT)methylation,isocitrate dehydrogenase(IDH)mutation and chromosome 1p19q co-deletion.The expression levels of AURKA mRNA in different histopathology types,WHO grades,and primary/recurrent status gliomas were analyzed.We compared the OS of glioma patients with different expression levels of AURKA mRNA using Kaplan-Meier survival curves.The effects of AURKA mRNA expression level,IDH mutation,chromosome 1p19q co-deletion,age,radiotherapy,chemotherapy,WHO grade and primary/recurrent status on the OS of glioma patients were assessed through univariate and multivariate Cox regression analyses.Receiver operating characteristic(ROC)curve was plotted and area under the curve(AUC)was used to evaluate the performance of AURKA mRNA expression level in predicting the OS of glioma patients.Tissue samples of gliomas were collected from patients undergoing surgical treatment at the Neurosurgery Center of Beijing Tiantan Hospital,Capital Medical University,including 4 low-grade gliomas(LCGG)and 4 GBM tissue samples.The expression of AURKA in the tumor tissue samples was detected by Western blot(WB).The human GBM cell line U87-MG was treated with AURKA inhibitor alisertib and U87-MG cells were divided into a control group(treated with DMSO)and an alisertib-treated group(treated with 5μmol/L alisertib).The effects of alisertib on GBM cells proliferation and colony formation were detected by cell counting Kit-8(CCK-8)test and crystal violet staining,respectively.The effects of alisertib on the expression of AURKA,phosphorylation Thr288 of AURKA and B7-H3 were detected by W

关 键 词：胶质母细胞瘤 细胞增殖 极光激酶A 免疫检查点 B7-H3蛋白 

分 类 号：R739.4[医药卫生—肿瘤]