摘要
目的:探讨基因TRIM22在人脑胶质母细胞瘤中的表达及对增殖、侵袭和迁移等生物能力的影响。方法:通过TCGA数据库分析胶质母细胞瘤组织和正常脑组织中TRIM22表达水平,采用实时定量PCR(qRT-PCR)和Western blot两种方法分别检测胶质母细胞瘤细胞系U87和U251中TRIM22 m RNA和蛋白表达水平。应用Lipofectamine 3000将2种TRIM22-si RNA分别转染至U87及U251细胞中,再分别通过q RT-PCR和Western blot检测胶质母细胞瘤细胞中TRIM22敲低效率。应用CCK-8法、细胞平板克隆形成实验来评估TRIM22敲低后对于细胞增殖能力的影响;应用细胞划痕实验和Transwell实验检测TRIM22敲低后细胞侵袭和迁移能力的变化。应用Western blot检测TRIM22敲低对PI3K/AKT信号通路蛋白和上皮间质化标记蛋白(N-cadherin、Vimentin、E-cadherin)表达的影响。结果:在U87及U251细胞中敲低TRIM22后,细胞增殖、侵袭和迁移都明显减弱,p-AKT(S473)、N-cadherin、Vimentin表达水平则明显降低,E-cadherin表达水平明显升高。结论:TRIM22可能通过调节PI3K/AKT信号通路调控胶质母细胞瘤增殖、侵袭和迁移能力。
Objective：To explore the effect of TRIM22 on proliferation,invasion and migration in glioblastoma. Methods：The expression levels of TRIM22 were analyzed in TCGA database and were examined in two glioblastoma cell lines（U87 and U251 cells）using q RT-PCR and Western blot. Two independent TRIM22 siRNAs were transfected into U87 and U251 cells,and the effect of TRIM22 depletion was confirmed. CCK8 assay and colony formation assay were carried out to assess the effect of TRIM22 depletion on glioblastoma cell proliferation. Transwell assay and wound healing assay were also performed to investigate the role of TRIM22 depletion on glioblastoma cell invasion and migration. The expression of p-AKT and epithelial-mesenchymal transition（EMT）-associated proteins including N-cadherin,Vimentin,E-cadherin were examined using Western blot after TRIM22 depletion. Results：Inhibition of TRIM22 expression markedly reduced the proliferation,invasion and migration of glioblastoma cells. Correspondingly,the expression of p-AKT（S473）,N-cadherin and Vimentin were decreased,whereas the expression of E-cadherin was increased after TRIM22 depletion. Conclusion：TRIM22 is a crucial factor for regulating the proliferation,invasion and migration of glioblastoma cells through PI3K/AKT signaling.
作者
冯爽
陈正新
仇文进
蔡小敏
陆嘉诚
刘宁
王慧博
Feng Shuang;Chen Zhengxin;Qiu Wenjin;Cai Xiaomin;Lu Jiacheng;Liu Ning;Wang Huibo(Department of Neurosurgery, the First Affiliated Hospital of NMU, Nanjing 210029, China)
出处
《南京医科大学学报（自然科学版）》
CAS
CSCD
北大核心
2018年第6期728-733,共6页
Journal of Nanjing Medical University(Natural Sciences)
基金
国家自然科学基金(81772681)
江苏省科技厅基础研究计划(BK20160098)
