High throughput determination of TGFβ1/SMAD3 targets in A549 lung epithelial cells.
High throughput determination of TGFβ1/SMAD3 targets in A549 lung epithelial cells.
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DOI：
10.1371/journal.pone.0020319
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Kaminski N
中科院分区：
文献类型：
--
作者：
Zhang Y;Handley D;Kaplan T;Yu H;Bais AS;Richards T;Pandit KV;Zeng Q;Benos PV;Friedman N;Eickelberg O;Kaminski N
转化生长因子β1 （tgf - β1）在包括肺癌、肺动脉高压和肺纤维化在内的许多肺部疾病中发挥重要作用。tgf - β1激活信号转导级联，主要通过dna结合转录因子SMAD3介导细胞核内基因的转录调控。本研究的目的是鉴定肺上皮细胞中SMAD3结合靶点的全基因组图谱以及tgf - β1/SMAD3信号通路和网络的影响。我们将染色质免疫沉淀与人类启动子区微阵列（ChIP-on-chip）以及基因表达微阵列相结合，研究了tgf - β1/SMAD3通路在人A549肺泡上皮细胞中的全局转录调控。利用计算方法确定tgf - β1/SMAD3信号传导相关的分子通路和网络。在A549和人原代肺上皮细胞上，通过实时定量RT-PCR和电泳迁移量转移实验验证了SMAD3基因的表达和启动子的直接结合。已知的tgf - β1靶基因如SERPINE1、SMAD6、SMAD7、TGFB1和LTBP3在ChIP-on-chip和基因表达分析中都被发现，以及一些以前未被识别的靶基因如FOXA2。确认SMAD3结合FOXA2启动子并改变表达。结合ChIP-on-chip芯片和基因表达芯片的计算方法揭示了tgf - β1/SMAD3信号通路影响的多种靶分子途径。tgf - β1/SMAD3信号的全局靶点和分子通路和网络的鉴定，有助于更好地理解纤维化和癌变过程中上皮细胞表型的决定机制，以及tgf - β1对FOXA2的直接作用的发现。
Transforming growth factor beta 1 (TGFβ1) plays a major role in many lung diseases including lung cancer, pulmonary hypertension, and pulmonary fibrosis. TGFβ1 activates a signal transduction cascade that results in the transcriptional regulation of genes in the nucleus, primarily through the DNA-binding transcription factor SMAD3. The objective of this study is to identify genome-wide scale map of SMAD3 binding targets and the molecular pathways and networks affected by the TGFβ1/SMAD3 signaling in lung epithelial cells. We combined chromatin immunoprecipitation with human promoter region microarrays (ChIP-on-chip) along with gene expression microarrays to study global transcriptional regulation of the TGFβ1/SMAD3 pathway in human A549 alveolar epithelial cells. The molecular pathways and networks associated with TGFβ1/SMAD3 signaling were identified using computational approaches. Validation of selected target gene expression and direct binding of SMAD3 to promoters were performed by quantitative real time RT-PCR and electrophoretic mobility shift assay on A549 and human primary lung epithelial cells. Known TGFβ1 target genes such as SERPINE1, SMAD6, SMAD7, TGFB1 and LTBP3, were found in both ChIP-on-chip and gene expression analyses as well as some previously unrecognized targets such as FOXA2. SMAD3 binding of FOXA2 promoter and changed expression were confirmed. Computational approaches combining ChIP-on-chip and gene expression microarray revealed multiple target molecular pathways affected by the TGFβ1/SMAD3 signaling. Identification of global targets and molecular pathways and networks associated with TGFβ1/SMAD3 signaling allow for a better understanding of the mechanisms that determine epithelial cell phenotypes in fibrogenesis and carcinogenesis as does the discovery of the direct effect of TGFβ1 on FOXA2.
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影响因子：
6.7
作者：
Latella, Giovanni;Vetuschi, Antonella;Gaudio, Eugenio
通讯作者：
Gaudio, Eugenio
DOI：
10.1111/j.1365-2362.2008.02076.x
发表时间：
2009-02-01
影响因子：
5.5
作者：
Latella, G.;Vetuschi, A.;Gaudio, E.
通讯作者：
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DOI：
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发表时间：
1999-01-01
影响因子：
1.6
作者：
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通讯作者：
Bonewald, LF
DOI：
10.1097/jto.0b013e3181ce3afd
发表时间：
2010-04
期刊：
Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer
影响因子：
--
作者：
Jeon HS;Jen J
通讯作者：
Jen J
影响因子：
4.4
作者：
Bonniaud, P;Kolb, M;Gauldie, J
通讯作者：
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