胶质瘤细胞中ADAR2 mRNA前体选择性模式的分析
首发时间:2013-12-27
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别。方法:RT-PCR产物测序检测胶质瘤细胞和正常胶质细胞中Glu2 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,Real-time 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胶质细胞中ADAR2 mRNA前体剪接模式的差异。结果:胶质瘤细胞中Glu2 Q/R位点的A-to-I编辑水平明显下降。Real-time PCR检测到在胶质瘤细胞中,Exon 5a(+)/5a(-)的比值明显高于正常星形胶质细胞HA1800。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差异。结论:Exon 5a位点的剪接在胶质瘤细胞和正常细胞中存在明显差别,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。
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Analysis the alternative splicing pattern of ADAR2 pre-mRNA in human glioma cell lines
Abstract:Objective:To analysis the differences of ADAR2 alternatively splicing pattern between glioma cell lines and normal human astrocyte. Methods:Real-time PCR was performed to detect the expression level of each alternatively splicing variant using specific primer which was confirmed to amplify only the targeted template and not other alternatively spliced variant fragments. Results:Real-time PCR revealed that the ADAR2 pre-mRNA splicing pattern has no significant difference at exon 1a and exon 2 between glioma cell lines and normal human astrocyte; At the same time, we 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 alternative splicing variants not including exon 5a in human astrocyte. Conclusion:The different alternatively splicing pattern at exon 5a may attribute to the decreased activity of ADAR2..
Keywords: Surgery Glioma ADAR2 Alternative splice
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胶质瘤细胞中ADAR2 mRNA前体选择性模式的分析
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