Abstract
Purpose
Epithelial mesenchymal transition (EMT) plays a central role in the development of fibrotic complications of the lens. The current study is designed to check whether EMT of lens epithelial cells (LECs) is regulated by epigenetic modifications and to evaluate the effect of Trichostatin-A (TSA) on the transforming growth factor-β (TGF-β)-induced EMT.
Methods
Fetal human LECs (FHL124) were treated with TGF-β2 in the presence or absence of TSA. Levels of mRNA, protein, as well as localization of α-smooth muscle actin (αSMA) were studied along with migration of LECs. Acetylation of histone H4 was analyzed and chromatin immunoprecipitation (ChIP) was carried out to study the level of acetylated histone H4 at the promoter of αSMA gene (ACTA2). Student’s t-test was used for statistical analysis.
Results
TGF-β2 treatment resulted in myofibroblast-like changes and increased migratory capacity of FHL124. Protein and mRNA expression of αSMA increased, and immunofluorescence revealed presence of extensive stress fibers. TSA treatment preserved epithelial morphology, retarded cell migration, and abrogated an increase in αSMA levels. TSA led to the accumulation of acetylated histone H4 that was reduced on TGF-β2 treatment. However, increased level of histone H4 acetylation was found at the ACTA2 promoter region during TGF-β treatment.
Conclusions
The increased level of αSMA, a hallmark of EMT in LECs, is associated with increased level of histone H4 acetylation at its promoter region, and TSA helps in suppressing EMT by epigenetically reducing this level. TSA thus shows promising potential in management of fibrotic conditions of the lens.
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Acknowledgements
This work was supported by WOS-A scheme of the Department of Science and Technology (SR/WOS-A/LS-272/2010). We thank Dr K Thangaraj of Centre for Cellular and Molecular Biology, Hyderabad, India, for his help with STR profiling. This paper was presented in part at the Asia-ARVO 2013 conference, New Delhi, India.
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Ganatra, D., Rajkumar, S., Patel, A. et al. Association of histone acetylation at the ACTA2 promoter region with epithelial mesenchymal transition of lens epithelial cells. Eye 29, 828–838 (2015). https://doi.org/10.1038/eye.2015.29
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DOI: https://doi.org/10.1038/eye.2015.29
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