Abstract
Alcohol affects approximately 1% (40,000) of new born infants each year and is the main preventable cause of mental retardation in the US. Ethanol alters cell signaling and promotes apoptosis and differentiation. Heparin-binding epidermal growth factor-like growth factor (HB-EGF), a member of the EGF family of growth factors, has been reported to prevent apoptosis and differentiation. We treated human embryonic stem cells (hESCs) with ethanol (20 mM) to reflect casual drinking, with and without HB-EGF to measure its ability to prevent ethanol-induced apoptosis and differentiation. Apoptosis was measured by DNA fragmentation (terminal dUTP nick-end labeling assays) and activated caspase-3, while differentiation was accessed by SSEA-1 and OCT-3/4; western blotting assessed MAPK signaling. HB-EGF reduced SSEA-1 and elevated OCT-3/4, while reducing the amount of activated caspase-3 and DNA fragmentation. Western blot analysis showed HB-EGF prevents ethanol from altering MAPK phosphorylation. This data suggests that ethanol-induced apoptosis was reduced by HB-EGF, while hESC pluripotency was maintained.
Acknowledgements
Special thanks to Dr Steven L. Stice, Director of the Regenerative Bioscience Center, The University of Georgia, Dr Gregory Buck, Texas A&M University-Corpus Christi, Dr Lou Ann S. Brown, Department of Pediatrics, Emory University. We thank Jagan Arumugham MS, for performing statistical analyses, Bethany Stewart BS for helping with tissue culture and Malini Krishnamoorthy MS, for her scientific insight. IANAFS. This work was supported by Emory University School of Medicine Department of Anesthesiology.
Declaration of interest: The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper.