摘要
目的 分析ANXA1 mRNA和蛋白在脑胶质母细胞瘤组织与正常脑组织之间的表达差异以及ANXA1在脑胶质母细胞瘤的表达,及其与脑胶质母细胞瘤患者预后的关系;体外探讨ANXA1对脑胶质母细胞瘤细胞系增殖、迁移、侵袭、凋亡能力的影响。方法 (1)从公共数据库获取脑胶质母细胞瘤组织及正常脑组织的表达数据及相应的临床数据。比较脑胶质母细胞瘤组织与正常脑组织之间ANXA1 mRNA的表达差异及ANXA1 mRNA高低表达组之间胶质母细胞瘤患者的生存时间差异。(2)通过人类蛋白质图谱公共数据库获取ANXA1蛋白在U251细胞内定位;免疫组化检测脑胶质母细胞瘤组织及其对应的癌旁组织中ANXA1蛋白的表达。(3)体外培养脑胶质母细胞瘤细胞系U87和U251,构建敲低ANXA1基因表达的小干扰RNA(ANXA1-siRNA)并转染至U87和U251细胞中。qRT-PCR和Western blot用于检测mRNA和蛋白的表达。细胞计数试剂盒(CCK-8)、Transwell和流式细胞术分别用于检测细胞增殖、细胞迁移和侵袭、细胞凋亡。结果 (1)在TCGA、Rembrandt和GSE16011队列中均发现ANXA1 mRNA在胶质母细胞瘤组织中表达明显上调,差异有统计学意义(P <0.05)。另外,在这三个队列中均发现ANXA1 mRNA高表达组患者总体生存时间明显低于低表达组患者(P <0.001)。(2)qRT-PCR和Western blot的分析结果均表明与阴性对照组相比,ANXA1-siRNA组ANXA1的表达水平明显降低,差异均有统计学意义(P <0.001)。(3)CCK-8检测结果表明敲低ANXA1表达后,U87和U251细胞的增殖能力明显低于阴性对照组,差异有统计学意义(P <0.001)。Transwell细胞迁移检测结果表明:敲低ANXA1表达后U87和U251细胞的迁移能力明显低于对照组[实验组:(25.40±2.70)、(11.20±4.44);阴性对照组:(55.20±8.70)、(32.20±8.11)];细胞侵袭能力明显低于对照组[实验组:(19.60±3.21)、(12.80±4.66);阴性对照组:(43.40±5.13)、(38.6±7.37)]能力均明显低于阴性对照组,
Objective To analyze the expression differences of ANXA1 mRNA and protein between glio⁃blastoma and normal brain tissue,and the relationship between ANXA1 expression in glioblastoma and prognosis of glioblastoma patients,and to investigate the effects of ANXA1 on the proliferation,migration,invasion and apoptosis of glioblastoma cell lines in vitro.Methods(1)The expression of glioblastoma tissues and normal brain tissues and corresponding clinical data were obtained from public databases.The difference of ANXA1 mRNA expression between glioblastoma tissues and normal brain tissues,and the difference of survival time of glioblastoma patients between high and low ANXA1 mRNA expression groups were compared.(2)The localization of ANXA1 protein in U251 cells was obtained from the Human Protein Atlas Public database.The expression of ANXA1 pro⁃tein in glioblastoma tissues and adjacent tissues was detected by immunohistochemistry.(3)Glioblastoma cell lines U87 and U251 were cultured in vitro,and ANXA1⁃siRNA knockdown ANXA1 gene expression was constructed and transferred into U87 and U251 cells.qRT⁃PCR and western blotting were used to detect mRNA and protein expression.Cell counting kit(CCK⁃8),Transwell and flow cytometry were used to detect cell proliferation,cell migration and invasion,and cell apoptosis,respectively.Results(1)ANXA1 mRNA expression was significantly up⁃regulated in glioblastoma tissues in TCGA,Rembrandt and GSE16011 cohorts,showing statistical significance(P<0.05).In addition,the overall survival time of patients with high ANXA1 mRNA expression was significantly lower than that of patients with low ANXA1 mRNA expression in all the three cohorts(P<0.001).(2)qRT⁃PCR and western blotting showed that compared with that in the negative control group,the expression level of ANXA1 in the ANXA1⁃siRNA group was significantly decreased,and the difference was statistically significant(P<0.001).(3)CCK⁃8 detection results showed that after knockdown of ANXA1 expression,the proliferation ability of
作者
李梦欣
惠磊
汲乾坤
王树仁
李海明
金保哲
LI Mengxin;HUI Lei;JI Qiankun;WANG Shuren;LI Haiming;JIN Baozhe(Department of Neurosurgery,the First Affiliated Hospital of Xinxiang Medical University,Xinxiang 453000,China)
出处
《实用医学杂志》
CAS
北大核心
2023年第8期928-935,共8页
The Journal of Practical Medicine
基金
河南省科技攻关计划省部共建项目(编号:SBGJ2018057)
河南省卫健委联合共建项目(编号:LHGJ20190445)。
