Gene expression profiling indicate role of ER stress in miR-23a∼27a∼24-2 cluster induced apoptosis in HEK293T cells
Gene expression profiling indicate role of ER stress in miR-23a∼27a∼24-2 cluster induced apoptosis in HEK293T cells
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DOI：
10.4161/rna.8.4.15583
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发表时间：
2011-07-01
期刊：
影响因子：
4.1
通讯作者：
Saini, Neeru
中科院分区：
文献类型：
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作者：
Chhabra, Ravindresh;Dubey, Richa;Saini, Neeru
在此之前，我们已经报道过类似于24-2簇的miR-23 a过表达通过JNK诱导人胚肾（HEK 293 T）细胞凋亡。在此，我们描述了负责miR-23 a类似于27 a类似于24-2簇诱导的细胞凋亡的分子机制。使用基因表达谱来表征在HE K293 T细胞中对miR-23 a（类似于27 a）（类似于24-2簇过表达）的转录应答。微阵列分析给出了1，025个差异表达的基因，并且使用Influence Pathway Analysis（IPA）软件对基因表达数据进行分析，揭示了受影响的顶级过程中的p53信号传导，氧化应激反应和线粒体功能障碍。这些数据证实了我们以前的研究，我们已经报道了ROS的增加和促凋亡因子，如细胞色素c（cyt c）和凋亡诱导因子（AIF）从线粒体的释放。此外，ER应激介导的细胞凋亡途径的组分，即，C/EBP同源蛋白（CHOP/DDIT 3/GADD 153）和Akt抑制剂TRIB 3在细胞内明显富集。此外，在miR-23 a中，类似于27 a，类似于24-2簇过表达的HE K293 T细胞，在RNA和蛋白质水平上观察到ATF 3和ATF 4的表达增强。BIM的诱导似乎是特异性的，因为ER应激仅引起相关BH 3-only蛋白BID或BH 3-only蛋白A表达的微小变化。miR-23 a类似于27 a类似于24-2簇触发内质网应激（ER应激）的事实通过在HE K293 T细胞中过表达该簇后细胞溶质钙水平的增加进一步确立。这些发现也支持PANTHER分析，其中细胞凋亡和应激反应的生物过程类别丰富。总之，这项工作强调了ER应激在miR-23 a（类似于27 a）（类似于24-2簇）介导的HE K293 T细胞凋亡中的作用。由于该簇诱导的细胞凋亡的详细知识现已阐明，该簇的体内研究将有助于评估其作为已知由于细胞凋亡失调而发生的疾病的治疗剂的用途的前景。
Previously, we had reported that overexpression of miR-23a similar to 27a similar to 24-2 cluster induces caspase-dependent and -independent apoptosis via JNK in human embryonic kidney (HEK293T) cells. Herein, we describe the molecular mechanism(s) responsible for miR-23a similar to 27a similar to 24-2 cluster induced apoptosis. Gene expression profiling was used to characterize the transcriptional response to miR-23a similar to 27a similar to 24-2 cluster overexpression in HE K293T cells. The microarray analysis gave 1,025 differentially expressed genes and analysis of the gene expression data with Ingenuity Pathway Analysis (IPA) software revealed p53 signaling, oxidative stress response and mitochondrial dysfunction among the top processes being affected. This data substantiates our previous study where we had reported increase of ROS and the release of proapoptotic factors such as cytochrome c (cyt c) and apoptosis-inducing factor (AIF) from the mitochondria. Additionally, components of ER stress-mediated apoptosis pathway i.e., C/EBP homologous protein (CHOP/DDIT3/GADD153) and TRIB3, an Akt inhibitor were found to be significantly enriched. Also, the enhanced expression of ATF3 and ATF4 was observed at RNA as well as protein level in miR-23a similar to 27a similar to 24-2 cluster overexpressed HE K293T cells. Induction of BIM appeared to be specific, because ER stress caused only a minor change in the expression of the related BH3-only proteins BID or PUMA. The fact that miR-23a similar to 27a similar to 24-2 cluster triggered endoplasmic reticulum stress (ER stress) was further established by the increase in cytosolic calcium levels after overexpression of this cluster in HE K293T cells. These findings were also supported by PANTHER analysis wherein biological process categories of apoptosis and stress response were enriched. Taken together, this work underlines the role of ER stress in miR-23a similar to 27a similar to 24-2 cluster mediated apoptosis in HE K293T cells. Since the detailed knowledge of this cluster induced apoptosis has now been elucidated, the in vivo study of this cluster would help evaluate the prospect of its use as a therapeutic in diseases known to occur because of deregulation of apoptosis.