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Thyroid hormone responsive genes in cultured human fibroblasts
Lars C Moeller1, Alexandra M Dumitrescu, Robert L Walker
1Department of Medicine, The University of Chicago, Illinois 60637, USA.
The Journal of Clinical Endocrinology and Metabolism
|October 28, 2004
Summary
Thyroid hormone (TH) influences gene expression in human skin fibroblasts, impacting over 90 genes. This study identifies new TH-responsive genes and confirms fibroblasts
Area of Science:
- Endocrinology
- Molecular Biology
- Genetics
Background:
- Human skin fibroblasts are accessible for studying inherited diseases and pathophysiology.
- Thyroid hormone (TH) plays a crucial role in cellular metabolism and development.
- Understanding TH's effect on gene expression is vital for comprehending its physiological roles.
Purpose of the Study:
- To investigate the impact of thyroid hormone (TH) on gene expression in normal human skin fibroblasts.
- To identify novel TH-responsive genes using quantitative fluorescent cDNA microarray analysis.
- To validate the specificity of TH's effects by comparing normal fibroblasts with those from patients with resistance to thyroid hormone (RTH).
Main Methods:
- Quantitative fluorescent cDNA microarray analysis of over 15,000 genes in normal human skin fibroblasts treated with TH.
- Comparison of gene expression profiles in normal fibroblasts versus fibroblasts from RTH patients with TH receptor-beta gene mutations.
- Validation of selected gene expression changes using real-time PCR and dose-response studies.
Main Results:
- Microarray analysis identified 91 up-regulated and five down-regulated genes in response to TH treatment (2 x 10(-9) M T(3) for 24 h).
- Real-time PCR confirmed the expression changes for 8 out of 10 induced and 2 out of 3 repressed genes.
- Several novel TH-responsive genes were identified, including aldo-keto reductase family 1 C1-3, collagen type VI alpha 3, and hypoxia-inducible factor-1 alpha.
- TH-mediated down-regulation of fibroblast growth factor 7 and alcohol dehydrogenase 1B was observed even in RTH patient fibroblasts.
Conclusions:
- Human skin fibroblasts are a suitable model for studying TH action in both normal physiology and disease states.
- This study identified several new TH-responsive genes involved in diverse regulatory functions, expanding our understanding of TH's molecular mechanisms.
- TH appears to regulate gene expression through both genomic and nongenomic pathways, with some effects persisting independently of TH receptor defects.