High throughput determination of TGFß1/SMAD3 targets in A549 lung epithelial cells.
PLoS One
; 6(5): e20319, 2011.
Article
em En
| MEDLINE
| ID: mdl-21625455
ABSTRACT
BACKGROUND:
Transforming growth factor beta 1 (TGFß1) plays a major role in many lung diseases including lung cancer, pulmonary hypertension, and pulmonary fibrosis. TGFß1 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 TGFß1/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 TGFß1/SMAD3 pathway in human A549 alveolar epithelial cells. The molecular pathways and networks associated with TGFß1/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 ANDCONCLUSIONS:
Known TGFß1 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 TGFß1/SMAD3 signaling. Identification of global targets and molecular pathways and networks associated with TGFß1/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 TGFß1 on FOXA2.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Proteína Smad3
/
Fator de Crescimento Transformador beta1
/
Pulmão
Tipo de estudo:
Prognostic_studies
Limite:
Humans
Idioma:
En
Revista:
PLoS One
Assunto da revista:
CIENCIA
/
MEDICINA
Ano de publicação:
2011
Tipo de documento:
Article
País de afiliação:
Estados Unidos