Document Detail


Human aldehyde dehydrogenase 3A1 inhibits proliferation and promotes survival of human corneal epithelial cells.
MedLine Citation:
PMID:  15905174     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Aldehyde dehydrogenase 3A1 (ALDH3A1) is a NAD(P)+-dependent enzyme that is highly expressed in mammalian corneal epithelial cells and has been shown to protect against UV- and 4-hydroxynonenal-induced cellular damage, mainly by metabolizing toxic lipid peroxidation aldehydes. Here we report a novel function of ALDH3A1 as a negative cell cycle regulator. We noticed a reduction in ALDH3A1 gene expression in actively proliferating primary human corneal epithelium explant cultures, indicating that ALDH3A1 expression is inversely correlated with replication. To examine this further, a human corneal epithelial cell line (HCE) lacking endogenous ALDH3A1 was stably transfected to express ALDH3A1 at levels similar to those found in vivo. ALDH3A1-transfected cells exhibited an elongated cell cycle, decreased plating efficiency, and reduced DNA synthesis compared with the mock-transfected cells. These effects were associated with reduced cyclin A- and cyclin B-dependent kinase activities and reduced phosphorylation of the retinoblastoma protein (pRb) as well as decreased protein levels of cyclins A, B, and E, the transcription factor E2F1, and the cyclin-dependent kinase inhibitor p21. These observations were further expanded and confirmed on human keratinocyte cells (NCTC-2544) overexpressing ALDH3A1. Consistent with a protective role of an elongated cell cycle, ALDH3A1-transfected cells exhibited increased resistance to the cytotoxic effects of the DNA-damaging agents mitomycin C and Vp-16. Immunohistochemistry and biochemical fractionation revealed that ALDH3A1 is localized both in the nucleus and cytosol of ALDH3A1-transfected cells, implying a possible association between the nuclear localization of the enzyme and its proliferation-suppressive functions. In conclusion, these results suggest that ALDH3A1 may protect corneal epithelial cells against oxidative damage not only through its metabolic function but also by prolonging the cell cycle.
Authors:
Aglaia Pappa; Donald Brown; Yiannis Koutalos; James DeGregori; Carl White; Vasilis Vasiliou
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.     Date:  2005-05-19
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  280     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  2005 Jul 
Date Detail:
Created Date:  2005-07-25     Completed Date:  2005-09-22     Revised Date:  2011-09-22    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  27998-8006     Citation Subset:  IM    
Affiliation:
Molecular Toxicology and Environmental Health Sciences Program, University Colorado Health Sciences Center, Denver, Colorado 80262, USA.
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MeSH Terms
Descriptor/Qualifier:
Aldehyde Dehydrogenase / metabolism,  physiology*
Aldehydes / pharmacology
Apoptosis
Blotting, Western
Bromodeoxyuridine / pharmacology
Cell Cycle
Cell Cycle Proteins / metabolism
Cell Nucleus / metabolism
Cell Proliferation
Cell Survival
Cells, Cultured
Cornea / cytology,  metabolism
Cyclin A / metabolism
Cyclin B / metabolism
Cyclin E / metabolism
Cytosol / metabolism
DNA / metabolism
DNA Damage
DNA Fragmentation
DNA-Binding Proteins / metabolism
E2F Transcription Factors
E2F1 Transcription Factor
Epithelial Cells / cytology*
Gene Expression Regulation
Humans
Immunohistochemistry
Keratinocytes / cytology
Microscopy, Confocal
Microscopy, Fluorescence
Mitomycin / pharmacology
Oxidative Stress
Protein Kinases / metabolism
Retinoblastoma Protein / metabolism
Time Factors
Transcription Factors / metabolism
Transfection
Ultraviolet Rays
Grant Support
ID/Acronym/Agency:
EY11490/EY/NEI NIH HHS; R01 CA077314-07/CA/NCI NIH HHS; T32 AA07464/AA/NIAAA NIH HHS
Chemical
Reg. No./Substance:
0/Aldehydes; 0/Cell Cycle Proteins; 0/Cyclin A; 0/Cyclin B; 0/Cyclin E; 0/DNA-Binding Proteins; 0/E2F Transcription Factors; 0/E2F1 Transcription Factor; 0/E2F1 protein, human; 0/Retinoblastoma Protein; 0/Transcription Factors; 29343-52-0/4-hydroxy-2-nonenal; 50-07-7/Mitomycin; 59-14-3/Bromodeoxyuridine; 9007-49-2/DNA; EC 1.2.1.3/ALDH3A1 protein, human; EC 1.2.1.3/Aldehyde Dehydrogenase; EC 2.7.-/Protein Kinases; EC 2.7.1.-/histone H1 kinase

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