贵州医科大学学报

2018, v.43;No.210(03) 269-273

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复制状态下EB病毒对宿主细胞中ZEB1基因表达的影响
Epstein-Barr Virus on ZEB1 Gene Expression in Host Cells under Replication

李明阳;马春霞;陈香岭;吴翰欣;吴小海;俞建昆;
LI Mingyang;MA Chunxia;CHEN Xiangling;WU Hanxin;WU Xiaohai;YU Jiankun;Institute of Medical Biology,Chinese Academy of Medical Sciences & Peking Union Medical College;Second Affiliated Hospital of Kunming Medical University;

摘要(Abstract):

目的:探究EB病毒(EBV)在潜伏感染和裂解复制状态下对ZEB1、ZEB2、ZEB1-AS1基因表达的影响。方法:将EBV相关癌细胞系Raji、SNU-719、AGS分为实验组(TPA及Na B诱导EBV)和对照组(加入等体积DMSO),将稳定表达p LVshRNA-EGFP-EBVsponge的SNU-719细胞系设为sponge组,未经转染RNA sponge的SNU-719细胞系设为control组(2组细胞均使用TPA及Na B进行诱导),TPA及Na B处理48 h后提取细胞总RNA进行RT-PCR,从mRNA水平检测ZEB1、ZEB2及ZEB1-AS1基因的表达。结果:与对照组相比,EBV复制期间,在Raji、SNU-719、AGS细胞中ZEB1 mRNA表达量显著升高(P<0.05),ZEB1-AS1 mRNA的表达量仅在AGS细胞中显著降低(P<0.05),在SNU-719和AGS细胞中未检测到ZEB2的表达;稳定表达p LVshRNA-EGFPEBVsponge的SNU-719细胞比未转染RNA sponge的SNU-719细胞,ZEB1表达量明显降低(P<0.05)。结论:EBV依靠自身编码的miRNA在复制期间促进ZEB1的表达,同时也对ZEB1-AS1有抑制作用。
Objective: To investigate the effect of EBV on the expression of ZEB1,ZEB2 and ZEB1-AS1 genes under latent infection and lytic replication. Method: The EBV associated cancer cell line Raji,SNU-719 and AGS was divided into two groups: the experimental group( TPA and Na B induced EBV) and the control group( Added equal volume DMSO). SNU-719 cell lines stably expressing p LVshRNA-EGFP-EBVsponge were divided into sponge group,and SNU-719 cells without transfection of RNA sponge were set to control group( both groups were induced by TPA and Na B). After 48 h of TPA and Na B treatment,total RNA was extracted for RT-PCR. and the expression of ZEB1,ZEB2 and ZEB1-AS1 genes was detected at mRNA levels. Result: The expression of ZEB1 mRNA in Raji,SNU-719 and AGS cells was significantly higher than that in the control group( P < 0. 05). The expression of ZEB1-AS1 mRNA in AGS cells only decreased significantly( P < 0. 05),but no ZEB2 expression was detected in SNU-719 and AGS cells. The expression of ZEB1 in SNU-719 cells with stable expression of p LVshRNA-EGFP-EBVsponge was significantly lower than that in SNU-719 cells without RNA sponge( P < 0. 05). Conclusion: EBV relies on self-encoded miRNA to promote the expression of ZEB1 during replication,and also inhibits the expression of ZEB1-AS1.

关键词(KeyWords): EB病毒;潜伏感染;裂解复制;E盒结合锋指蛋白1;miRNA;miRNA抑制剂
EB virus;latent infection;cleavage replication;zinc finger E-box bindingprotein;miRNA;RNA sponge

Abstract:

Keywords:

基金项目(Foundation): 高等学校博士学科点专项科研基金项目(20111106120056);; 中国医学科学院重大协同创新项目(2016-12M-001)

作者(Author): 李明阳;马春霞;陈香岭;吴翰欣;吴小海;俞建昆;
LI Mingyang;MA Chunxia;CHEN Xiangling;WU Hanxin;WU Xiaohai;YU Jiankun;Institute of Medical Biology,Chinese Academy of Medical Sciences & Peking Union Medical College;Second Affiliated Hospital of Kunming Medical University;

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DOI: 10.19367/j.cnki.1000-2707.2018.03.005

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