TY - JOUR
T1 - Aldehyde dehydrogenase 3A1 confers oxidative stress resistance accompanied by altered DNA damage response in human corneal epithelial cells
AU - Voulgaridou, Georgia Persephoni
AU - Tsochantaridis, Ilias
AU - Tolkas, Christos
AU - Franco, Rodrigo
AU - Giatromanolaki, Alexandra
AU - Panayiotidis, Mihalis I.
AU - Pappa, Aglaia
N1 - Funding Information:
This research was supported by: i) the European Union (European Social Fund-ESF) and Greek national funds through the Operational Program “Education and Lifelong Learning” of the National Strategic Reference Framework (NSRF) − Research Funding Program: Heracleitus II . Investing in knowledge society through the European Social Fund (to G.P.V) and ii) a Bodossaki Foundation scholarship (to I.T.). Part of this work was implemented in the facilities of “OPENSCREEN-GR” (MIS 5002691), implemented under the Action “ Reinforcement of the Research and Innovation Infrastructure ”, funded by the Operational Programme “Competitiveness, Entrepreneurship and Innovation” ( NSRF 2014–2020 ) and co-financed by Greece and the European Union (European Regional Development Fund) .
Publisher Copyright:
© 2020 Elsevier Inc.
PY - 2020/4
Y1 - 2020/4
N2 - Aldehyde dehydrogenase 3A1 is constitutively expressed in a taxon-specific manner in the cornea, where, due to its high abundance, it has been characterized as a corneal crystallin. ALDH3A1 has been proposed to be a multifaceted protein that protects cellular homeostasis through several modes of action. The present study examines the mechanisms by which ALDH3A1 exerts its cytoprotective role under conditions of oxidative stress. To this end, we have utilized an isogenic HCE-2 (human corneal epithelium) cell line pair differing in the expression of ALDH3A1. Single cell gel electrophoresis assay and H2DCFDA analysis revealed that the expression of ALDH3A1 protected HCE-2 cells from H2O2-, tert-butyl peroxide- and etoposide-induced oxidative and genotoxic effects. Furthermore, comparative qPCR analysis revealed that a panel of cell cycle (Cyclins B1, B2, D, E), apoptosis (p53, BAX, BCL-2, BCL-XL) and DNA damage response (DNA-PK, NBS1) genes were up-regulated in the ALDH3A1 expressing HCE-2 cells. Moreover, the expression profile of a variety of DNA damage signaling (DDS)-related genes, was investigated (under normal and oxidative stress conditions) by utilizing the RT2 profiler™ PCR array in both isogenic HCE-2 cell lines. Our results demonstrated that several genes associated with ATM/ATR signaling, cell cycle regulation, apoptosis and DNA damage repair were differentially expressed under all conditions tested. In conclusion, this study suggests that ALDH3A1 significantly contributes to the antioxidant defense of corneal homeostasis by maintaining DNA integrity possibly through altering the expression of specific DDS-related genes. Further studies will shed light on the precise role(s) of this multifunctional protein.
AB - Aldehyde dehydrogenase 3A1 is constitutively expressed in a taxon-specific manner in the cornea, where, due to its high abundance, it has been characterized as a corneal crystallin. ALDH3A1 has been proposed to be a multifaceted protein that protects cellular homeostasis through several modes of action. The present study examines the mechanisms by which ALDH3A1 exerts its cytoprotective role under conditions of oxidative stress. To this end, we have utilized an isogenic HCE-2 (human corneal epithelium) cell line pair differing in the expression of ALDH3A1. Single cell gel electrophoresis assay and H2DCFDA analysis revealed that the expression of ALDH3A1 protected HCE-2 cells from H2O2-, tert-butyl peroxide- and etoposide-induced oxidative and genotoxic effects. Furthermore, comparative qPCR analysis revealed that a panel of cell cycle (Cyclins B1, B2, D, E), apoptosis (p53, BAX, BCL-2, BCL-XL) and DNA damage response (DNA-PK, NBS1) genes were up-regulated in the ALDH3A1 expressing HCE-2 cells. Moreover, the expression profile of a variety of DNA damage signaling (DDS)-related genes, was investigated (under normal and oxidative stress conditions) by utilizing the RT2 profiler™ PCR array in both isogenic HCE-2 cell lines. Our results demonstrated that several genes associated with ATM/ATR signaling, cell cycle regulation, apoptosis and DNA damage repair were differentially expressed under all conditions tested. In conclusion, this study suggests that ALDH3A1 significantly contributes to the antioxidant defense of corneal homeostasis by maintaining DNA integrity possibly through altering the expression of specific DDS-related genes. Further studies will shed light on the precise role(s) of this multifunctional protein.
KW - ALDH3A1
KW - ALDHs
KW - Aldehyde dehydrogenase 3A1
KW - Antioxidant
KW - Corneal homeostasis
KW - DDR
KW - DDS
KW - DNA damage
KW - DNA damage response
KW - DNA damage signaling
KW - Oxidative stress
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UR - http://www.scopus.com/inward/citedby.url?scp=85079644138&partnerID=8YFLogxK
U2 - 10.1016/j.freeradbiomed.2020.01.183
DO - 10.1016/j.freeradbiomed.2020.01.183
M3 - Article
C2 - 32006654
AN - SCOPUS:85079644138
SN - 0891-5849
VL - 150
SP - 66
EP - 74
JO - Free Radical Biology and Medicine
JF - Free Radical Biology and Medicine
ER -