Zhang, Y and Handley, D and Kaplan, T and Yu, H and Bais, AS and Richards, T and Pandit, KV and Zeng, Q and Benos, PV and Friedman, N and Eickelberg, O and Kaminski, N
(2011)
High throughput determination of TGFβ1/SMAD3 targets in A549 lung epithelial cells.
PLoS ONE, 6 (5).
Abstract
Background: 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. Methodology: 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. Results and Conclusions: 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. © 2011 Zhang et al.
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Item Type: |
Article
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Status: |
Published |
Creators/Authors: |
Creators | Email | Pitt Username | ORCID ![](/images/orcid_id_24x24.png) |
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Zhang, Y | zhang3@pitt.edu | ZHANG3 | | Handley, D | | | | Kaplan, T | | | | Yu, H | | | | Bais, AS | | | | Richards, T | | | | Pandit, KV | | | | Zeng, Q | qiz30@pitt.edu | QIZ30 | | Benos, PV | benos@pitt.edu | GSBCPLRC | | Friedman, N | | | | Eickelberg, O | | | | Kaminski, N | | | |
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Contributors: |
Contribution | Contributors Name | Email | Pitt Username | ORCID ![](/images/orcid_id_24x24.png) |
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Editor | Rattray, Magnus | UNSPECIFIED | UNSPECIFIED | UNSPECIFIED |
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Date: |
26 May 2011 |
Date Type: |
Publication |
Journal or Publication Title: |
PLoS ONE |
Volume: |
6 |
Number: |
5 |
DOI or Unique Handle: |
10.1371/journal.pone.0020319 |
Schools and Programs: |
School of Medicine > Critical Care Medicine |
Refereed: |
Yes |
MeSH Headings: |
Base Sequence; Cell Line; Chromatin Immunoprecipitation; DNA Primers; Electrophoretic Mobility Shift Assay; Epithelial Cells--metabolism; Humans; Lung--cytology; Lung--metabolism; Oligonucleotide Array Sequence Analysis; Promoter Regions, Genetic; Protein Binding; Reverse Transcriptase Polymerase Chain Reaction; Smad3 Protein--metabolism; Transforming Growth Factor beta1--metabolism |
Other ID: |
NLM PMC3098871 |
PubMed Central ID: |
PMC3098871 |
PubMed ID: |
21625455 |
Date Deposited: |
29 Aug 2012 21:08 |
Last Modified: |
29 Oct 2022 11:55 |
URI: |
http://d-scholarship.pitt.edu/id/eprint/13858 |
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