Babenko Vladimir N, Gubanova Natalya V, Bragin Anatoly O, Chadaeva Irina V, Vasiliev Gennady V, Medvedeva Irina V, Gaytan Alexey S, Krivoshapkin Alexey L, Orlov Yuriy L
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J Integr Bioinform. 2017 Sep 18;14(3):20170022. doi: 10.1515/jib-2017-0022.
Here we present the analysis of alternative splicing events on an example of glioblastoma cell culture samples using a set of computer tools in combination with database integration. The gene expression profiles of glioblastoma were obtained from cell culture samples of primary glioblastoma which were isolated and processed for RNA extraction. Transcriptome profiling of normal brain samples and glioblastoma were done by Illumina sequencing. The significant differentially expressed exon-level probes and their corresponding genes were identified using a combination of the splicing index method. Previous studies indicated that tumor-specific alternative splicing is important in the regulation of gene expression and corresponding protein functions during cancer development. Multiple alternative splicing transcripts have been identified as progression markers, including generalized splicing abnormalities and tumor- and stage-specific events. We used a set of computer tools which were recently applied to analysis of gene expression in laboratory animals to study differential splicing events. We found 69 transcripts that are differentially alternatively spliced. Three cancer-associated genes were considered in detail, in particular: APP (amyloid beta precursor protein), CASC4 (cancer susceptibility candidate 4) and TP53. Such alternative splicing opens new perspectives for cancer research.
在此,我们以胶质母细胞瘤细胞培养样本为例,结合数据库整合,运用一系列计算机工具对可变剪接事件进行分析。胶质母细胞瘤的基因表达谱取自原发性胶质母细胞瘤的细胞培养样本,这些样本经分离后进行RNA提取处理。正常脑样本和胶质母细胞瘤的转录组分析通过Illumina测序完成。运用剪接指数法组合,鉴定出显著差异表达的外显子水平探针及其相应基因。先前的研究表明,肿瘤特异性可变剪接在癌症发展过程中对基因表达及相应蛋白质功能的调控中起着重要作用。多种可变剪接转录本已被确定为进展标志物,包括普遍的剪接异常以及肿瘤和阶段特异性事件。我们使用了一组最近应用于实验动物基因表达分析的计算机工具来研究差异剪接事件。我们发现了69个发生差异可变剪接的转录本。详细研究了三个与癌症相关的基因,具体为:APP(淀粉样前体蛋白)、CASC4(癌症易感性候选基因4)和TP53。这种可变剪接为癌症研究开辟了新的视角。