Transcriptome profiling reveals novel gene expression signatures and regulating transcription factors of TGFβ-induced epithelial-to-mesenchymal transition.
Transcriptome profiling reveals novel gene expression signatures and regulating transcription factors of TGFβ-induced epithelial-to-mesenchymal transition.
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DOI：
10.1002/cam4.719
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发表时间：
2016-08
期刊：
影响因子：
4
通讯作者：
Kuo MD
中科院分区：
文献类型：
--
作者：
Du L;Yamamoto S;Burnette BL;Huang D;Gao K;Jamshidi N;Kuo MD
癌细胞上皮向间充质转化(EMT)的失调性赋予癌细胞侵袭性和转移性，有利于肿瘤细胞在远端靶器官的成功定植，从而在早期癌向侵袭性恶性肿瘤的转化中发挥关键作用。EMT还与肿瘤复发、耐药和肿瘤干细胞启动有关。因此，更好地了解EMT背后的机制最终可能有助于开发专门针对EMT过程的新的预后方法和个体化治疗。为了研究EMT过程中中枢转录组的变化，我们建立了一个基于转化生长因子-β的体外EMT模型，并利用该模型分析了两个不同的细胞系，非小细胞肺癌细胞系H358和乳腺细胞系MCF10A中与EMT相关的基因转录变化。经转化生长因子-β/肿瘤抑素M(OSM)处理7天后，观察到细胞形态转变为间充质表型，以及与EMT相关的mRNA和蛋白表达的变化。此外，注意到移动性增加，流式细胞术证实肿瘤干细胞样群中有丰富的细胞。基于微阵列的差异表达分析发现了一个与EMT相关的基因表达特征，该特征被RT-qPCR证实，并与先前发表的EMT核心特征显著重叠。最后，鉴定并独立验证了两个新的EMT调控转录因子IRF5和LMCD1。
Dysregulated epithelial to mesenchymal transition (EMT) in cancer cells endows invasive and metastatic properties upon cancer cells that favor successful colonization of distal target organs and therefore play a critical role in transforming early‐stage carcinomas into invasive malignancies. EMT has also been associated with tumor recurrence and drug resistance and cancer stem cell initiation. Therefore, better understanding of the mechanisms behind EMT could ultimately contribute to the development of novel prognostic approaches and individualized therapies that specifically target EMT processes. As an effort to characterize the central transcriptome changes during EMT, we have developed a Transforming growth factor (TGF)‐beta‐based in vitro EMT model and used it to profile EMT‐related gene transcriptional changes in two different cell lines, a non‐small cell lung cancer cell line H358, and a breast cell line MCF10a. After 7 days of TGF‐beta/Oncostatin M (OSM) treatment, changes in cell morphology to a mesenchymal phenotype were observed as well as concordant EMT‐associated changes in mRNA and protein expression. Further, increased motility was noted and flow cytometry confirmed enrichment in cancer stem cell‐like populations. Microarray‐based differential expression analysis identified an EMT‐associated gene expression signature which was confirmed by RT‐qPCR and which significantly overlapped with a previously published EMT core signature. Finally, two novel EMT‐regulating transcription factors, IRF5 and LMCD1, were identified and independently validated.