Deep sequencing and in silico analyses identify MYB-regulated gene networks and signaling pathways in pancreatic cancer.
Deep sequencing and in silico analyses identify MYB-regulated gene networks and signaling pathways in pancreatic cancer.
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DOI：
10.1038/srep28446
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发表时间：
2016-06-29
影响因子：
4.6
通讯作者：
Singh AP
中科院分区：
文献类型：
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作者：
Azim S;Zubair H;Srivastava SK;Bhardwaj A;Zubair A;Ahmad A;Singh S;Khushman M;Singh AP
我们最近证实了转录因子MYB可以调节胰腺癌中几种癌症相关的表型。为了了解这些MYB相关变化的分子基础，我们对MYB过表达和沉默的胰腺癌细胞的转录组进行了深度测序，然后进行了计算机途径分析。我们鉴定了774个基因对MYB沉默的显著调节（p < 0.05），通过计算机分析将其分配到25个基因网络。进一步的分析将我们的RNA测序生成的数据集中的基因置于几个典型的信号传导途径，如细胞周期控制，DNA损伤和修复反应，p53和HIF 1 α。重要的是，我们观察到MYB沉默的胰腺癌细胞中胰腺癌信号通路的下调，表现出EGFR和NF-κB的抑制。通过qPCR和免疫印迹验证EGFR和RELA的表达降低，并且它们均显示在MYB的直接转录控制下。这些观察结果在匡威的方法中得到进一步证实，其中MYB在MYB缺失的胰腺癌细胞系中异位表达。因此，我们的研究结果表明，MYB可能通过靶向复杂的基因网络和信号通路调节胰腺癌细胞的生长和基因组稳定性。进一步深入的功能研究是必要的，以充分了解MYB信号在胰腺癌中的作用。
We have recently demonstrated that the transcription factor MYB can modulate several cancer-associated phenotypes in pancreatic cancer. In order to understand the molecular basis of these MYB-associated changes, we conducted deep-sequencing of transcriptome of MYB-overexpressing and -silenced pancreatic cancer cells, followed by in silico pathway analysis. We identified significant modulation of 774 genes upon MYB-silencing (p < 0.05) that were assigned to 25 gene networks by in silico analysis. Further analyses placed genes in our RNA sequencing-generated dataset to several canonical signalling pathways, such as cell-cycle control, DNA-damage and -repair responses, p53 and HIF1α. Importantly, we observed downregulation of the pancreatic adenocarcinoma signaling pathway in MYB-silenced pancreatic cancer cells exhibiting suppression of EGFR and NF-κB. Decreased expression of EGFR and RELA was validated by both qPCR and immunoblotting and they were both shown to be under direct transcriptional control of MYB. These observations were further confirmed in a converse approach wherein MYB was overexpressed ectopically in a MYB-null pancreatic cancer cell line. Our findings thus suggest that MYB potentially regulates growth and genomic stability of pancreatic cancer cells via targeting complex gene networks and signaling pathways. Further in-depth functional studies are warranted to fully understand MYB signaling in pancreatic cancer.