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深度测序揭示了喜树碱诱导凋亡过程中具有不同miRNA谱的两个Jurkat亚群。

Deep sequencing reveals two Jurkat subpopulations with distinct miRNA profiles during camptothecin-induced apoptosis.

作者信息

Erdoğan İpek, Coşacak Mehmet İlyas, Nalbant Ayten, Akgül Bünyamin

机构信息

Department of Molecular Biology and Genetics, İzmir Institute of Technology , Gülbahçeköyü, Urla, İzmir , Turkey.

出版信息

Turk J Biol. 2018 Apr 27;42(2):113-122. doi: 10.3906/biy-1710-62. eCollection 2018.

Abstract

MicroRNAs (miRNAs) are small noncoding RNAs of about 19-25 nt that regulate gene expression posttranscriptionally under various cellular conditions, including apoptosis. The miRNAs involved in modulation of apoptotic events in T cells are partially known. However, heterogeneity associated with cell lines makes it difficult to interpret gene expression signatures, especially in cancer-related cell lines. Treatment of the Jurkat T-cell leukemia cell line with the universal apoptotic drug, camptothecin, resulted in identification of two Jurkat subpopulations: one that is sensitive to camptothecin and another that is rather intrinsically resistant. We sorted apoptotic Jurkat cells from nonapoptotic ones prior to profiling miRNAs through deep sequencing. Our data showed that a total of 184 miRNAs were dysregulated. Interestingly, the apoptotic and nonapoptotic subpopulations exhibited distinct miRNA expression profiles. In particular, 6 miRNAs were inversely expressed in these two subpopulations. The pyrosequencing results were validated by real-time qPCR. Altogether, these results suggest that miRNAs modulate apoptotic events in T cells and that cellular heterogeneity requires careful interpretation of miRNA expression profiles obtained from drug-treated cell lines.

摘要

微小RNA(miRNA)是一类长度约为19 - 25个核苷酸的小型非编码RNA,在包括细胞凋亡在内的各种细胞条件下通过转录后水平调控基因表达。参与调节T细胞凋亡事件的miRNA已部分为人所知。然而,与细胞系相关的异质性使得解读基因表达特征变得困难,尤其是在癌症相关细胞系中。用通用凋亡药物喜树碱处理Jurkat T细胞白血病细胞系后,鉴定出两个Jurkat亚群:一个对喜树碱敏感,另一个具有内在抗性。在通过深度测序分析miRNA之前,我们将凋亡的Jurkat细胞与非凋亡细胞进行了分选。我们的数据显示共有184种miRNA表达失调。有趣的是,凋亡和非凋亡亚群表现出不同的miRNA表达谱。特别是,有6种miRNA在这两个亚群中呈反向表达。焦磷酸测序结果通过实时定量PCR得到验证。总之,这些结果表明miRNA调节T细胞中的凋亡事件,并且细胞异质性需要仔细解读从药物处理的细胞系中获得的miRNA表达谱。

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