Plk4在脑胶质瘤中的功能及其分子机制研究  

Study on the function and molecular mechanism of Plk4 in gliomas

作　　者：张晓阳 魏成 彭大钊 李生辉[1] 周俊虎[1] 梁浩[1] 韩磊[1] Zhang Xiaoyang;Wei Cheng;Peng Dazhao;Li Shenghui;Zhou Junhu;Liang Hao;Han Lei(Department of Neurosurgery,General Hospital of Tianjin Medical University,Tianjin Neurology Institute,Tianjin 300052,China)

机构地区：[1]天津医科大学总医院神经外科,天津市神经病学研究所,300052

出　　处：《中华神经外科杂志》2021年第11期1159-1166,共8页Chinese Journal of Neurosurgery

基　　金：国家自然科学基金(81773187,81572496)。

摘　　要：目的探讨Polo样激酶4(Plk4)在脑胶质瘤中的表达特征、生物学功能及其相关分子机制。方法纳入癌症基因组图谱(TCGA)数据库(689例)、中国胶质瘤基因组图谱(CGGA)数据库(693例)中脑胶质瘤患者的胶质瘤样本的转录组和临床数据, 纳入基因型和基因表达量关联(GTEx)数据库中正常脑组织标本的转录组数据(255例)。比较脑胶质瘤标本与正常脑组织中, 以及不同病理学分级脑胶质瘤标本中Plk4 mRNA表达量的差异;采用log-rank检验法比较Plk4 mRNA低表达组与高表达组患者总生存期的差异。取人星形胶质细胞、U87、LN229、U251及A172细胞株行相关研究。采用小干扰RNA(siRNA)转染法敲低U87和LN229细胞中Plk4的表达;采用实时荧光定量聚合酶链反应法检测Plk4 mRNA的表达;采用蛋白质免疫印迹法检测Plk4蛋白的表达;采用EdU实验和Transwell实验观察Plk4对胶质瘤细胞增殖、迁移及侵袭能力的影响。取4周龄雌性BALB/c-nu裸鼠20只, 将U87细胞注射于小鼠腹部制备胶质瘤异种荷瘤模型, 观察Plk4敲低组(肿瘤内注射siRNA-Plk4, n=10)与无义序列组(肿瘤内注射无序列Plk4, n=10)治疗21 d(7次)后肿瘤体积和重量的差异。取敲低Plk4的U87细胞, 采用蛋白组学和磷酸化组学的关联分析方法探讨Plk4在胶质瘤中的生物学功能及其分子机制。结果与星形胶质细胞相比, U87、LN229、U251及A172细胞株中Plk4 mRNA的表达水平均升高;TCGA联合GTEx数据库数据分析显示, 低级别胶质瘤及胶质母细胞瘤标本中Plk4 mRNA的表达量均高于正常脑组织;对CGGA数据库中标本的分析显示, Plk4 mRNA的表达量随世界卫生组织(WHO)病理学分级的升高而升高;2个数据库中Plk4 mRNA高表达组患者的总生存期均短于低表达组患者。EdU实验显示, 敲低Plk4表达的U87和LN229细胞的EdU阳性率均低于其无义序列细胞。Transwell实验显示, 敲低Plk4表达的U87和LN229细胞的穿胶数和迁移数�Objective To investigate the clinical characteristics,biological functions and molecular mechanism of polo-like Kinase 4(Plk4)in gliomas.Methods We studied the transcriptome data of glioma samples and clinical data of glioma patients in the Cancer Genome Atlas(TCGA)database(689 cases)and the Chinese Glioma Genome Atlas(CGGA)database(693 cases),and transcriptome data of nornial brain tissue specimens in the GIEx database(255 cases)were also included.The differences in the expression of Plk4 inglioma specitmens and normal brain tissues and glioma specimens of different pathological grades were compared.Log-rank lest was used to compare the differences in overall survival between patients with low Plk4 expressionand high expression groups.We took human astrocytes,U87,IN229,u251 and A172 cell lines for relatedresearch.Small interfering RNA(siRNA)transfection method was used to knock down the expression of Ple4 inU87 and LN229 cells.Real-time fluorescent quantitative polymerase chain reaction was used to detect theexpression of Plk4 mRNA.Western blotting was used to detect the expression of Plk4 protein.EU experimentand Transwell experiment were used to observe the effect of Plk4 on the proliferation,migration and invasion ofglioma cells.Twenty 4-week-old female BALB/c-mu nude mice were taken,and U87 cells were injected into theabdomen of mice to prepare glioma xenogeneic tumor-bearing models.The differences in tumor volume and weightwere analyzed between the Plk4 knockdown treatment group(intratumor injection of siRNA-Plk4)and the controlgroup(injection of non-sequence Plk4)after 21 d(7 times)of treatment.We took U87 cells with knocked downPlk4,and used proteomics and phosphorylation analysis to explore the molecular mechanism of Plk4s biologicalfunction in gliomas.Results Compared with astrocytes,the expression level of PLk4 mRNA in U87,LN229,U251 and Ai72 glioma cells was higher.TCGA combined with GTEx database data analysis showed that theexpression level of Plk4 mRNA in low-grade glioma and glioblastoma specimens

关 键 词：神经胶质瘤 蛋白激酶类 Polo样激酶4 蛋白组学 磷酸化组学 

分 类 号：R739.41[医药卫生—肿瘤]