胶质瘤细胞中ADAR2 mRNA前体选择性剪接模式的分析  

Analysis of alternative splicing pattern of ADAR2 pre-mRNA in human glioma cell lines

作　　者：李朝晖[1] 田男[2] 魏君[1] 李小玲[2] 杜超[1] 李妍哲 田宇[1] 

机构地区：[1]吉林大学白求恩医学部中日联谊医院神经外二科,长春市130031 [2] 浙江中医药大学生命科学学院

出　　处：《中国肿瘤临床》2014年第8期485-488,共4页Chinese Journal of Clinical Oncology

基　　金：吉林省科技发展计划国际科技合作项目(编号:20130413028GH)资助~~

摘　　要：目的:比较胶质瘤细胞U87、U251、A172和正常星形胶质细胞HA1800中ADAR2 mRNA前体选择性剪接模式的差异。方法:RFPCR产物测序检测胶质瘤细胞和正常胶质细胞中GluR2 Q/R位点的A-to-I编辑水平。根据前期研究鉴定出的选择性剪接位点设计特异性引物,RT-PCR方法检测胶质瘤细胞U87、U251、A172和正常星形胶质细胞HA1800中不同剪接转录本的相对表达量,比较胶质瘤细胞和正常星形胶质细胞中ADAR2 mRNA前体剪接模式的差异。结果:胶质瘤细胞中GluR2 Q/R位点的A-to-I编辑水平明显下降。RT-PCR检测到在胶质瘤细胞中,Exon 1a(+)/1a(-)、Exon 2(+)/2(-)的比值在胶质瘤细胞和正常星形胶质细胞中无显著性差异。Exon 5a(+)/5a(-)的比值明显高于正常星形胶质细胞HA1800。结论:Exon 5a位点的剪接在胶质瘤细胞和正常星形胶质细胞中有显著性差异,Exon 5a(+)转录本的表达增加可能是导致胶质瘤细胞中ADAR2编辑活性下降的原因。Objective：This study aims to analyze the differences in the alternative splicing pattern of ADAR2 among glioma cell lines U87, U251, A172, and normal human astrocyte HA1800. Methods：A-to-I editing level at the Q/R-Site of GluR-2 was analyzed by RT-PCR and sequencing. Real-time PCR was performed to detect the expression level of each alternatively splicing variant using a specific primer that was confirmed to amplify only the targeted template and not other alternatively spliced variant fragments. Results：We verified that the Q/R-Site of GluR-2 is under-edited in glioma cell lines. Real-time PCR revealed that the ADAR2 pre-mRNA splic-ing pattern has no significant difference at exons 1a and 2 between glioma cell lines and normal human astrocyte. We also detected that the amount of alternative splicing variants, including exon 5a, was higher than that of alternative splicing variants not including exon 5a in human glioma cell lines. However, the expression of alternative splicing variants, including exon 5a, was lower than that of alterna-tive splicing variants not including exon 5a in human astrocyte. Conclusion：Evident differences in splicing were observed at the site of exon 5a between glioma cell lines and normal human astrocytes. The difference in the alternatively splicing pattern at exon 5a may be attributed to the decreased activity of ADAR2.

关 键 词：胶质瘤 RNA编辑 选择性剪接 

分 类 号：R739.41[医药卫生—肿瘤]