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Induction and Analysis of Epithelial to Mesenchymal Transition
Published on: August 27, 2013
Proteomics analysis of epithelial cells reprogrammed in cell-free extract
Emma Pewsey1, Christine Bruce, A Stephen Georgiou
1Academic Unit of Reproductive and Developmental Medicine, University of Sheffield, Level 4, The Jessop Wing, Sheffield S102SF, United Kingdom.
Molecular & Cellular Proteomics : MCP
|March 3, 2009
Summary
Researchers reprogrammed epithelial cells to a pluripotent state using embryonal carcinoma cell extracts. This study reveals specific protein alterations during this nuclear reprogramming, offering insights for regenerative medicine.
Area of Science:
- Cell Biology
- Proteomics
- Regenerative Medicine
Background:
- Cellular reprogramming to pluripotency holds promise for regenerative medicine.
- Understanding the molecular mechanisms of reprogramming is crucial.
Purpose of the Study:
- To investigate the proteomic changes in epithelial cells reprogrammed to pluripotency.
- To characterize the protein profile of cells treated with embryonal carcinoma cellular extracts.
Main Methods:
- 293T epithelial cells were permeabilized and incubated with NCCIT cellular extracts.
- Gene expression (OCT4, SOX2, DNMT3B) and cell surface marker (SSEA-4) analysis were performed.
- Proteomic analysis using two-dimensional PAGE and tandem mass spectrometry was conducted.
Main Results:
- NCCIT extract treatment led to upregulation of OCT4 and SOX2 gene expression.
- 30% of treated cells expressed SSEA-4, confirming reprogramming.
- Proteomic analysis identified specific protein alterations, including 78-kDa glucose-regulated protein precursor and tropomyosin alpha-3 chain.
Conclusions:
- This study provides the first proteomic characterization of nuclear reprogramming.
- Specific protein alterations occur in epithelial cells reprogrammed by embryonal carcinoma extracts.
- These findings contribute to understanding cellular reprogramming for regenerative medicine applications.
