PMID- 33887608 OWN - NLM STAT- MEDLINE DCOM- 20210705 LR - 20240504 IS - 2213-2317 (Electronic) IS - 2213-2317 (Linking) VI - 43 DP - 2021 Jul TI - Activation of NRF2 by APE1/REF1 is redox-dependent in Barrett's related esophageal adenocarcinoma cells. PG - 101970 LID - S2213-2317(21)00118-X [pii] LID - 10.1016/j.redox.2021.101970 [doi] LID - 101970 AB - BACKGROUND: Chronic gastroesophageal reflux disease (GERD) is a major risk factor for the development of metaplastic Barrett's esophagus (BE) and its progression to esophageal adenocarcinoma (EAC). Uncontrolled accumulation of reactive oxygen species (ROS) in response to acidic bile salts (ABS) in reflux conditions can be lethal to cells. In this study, we investigated the role of APE1/REF1 in regulating nuclear erythroid factor-like 2 (NRF2), the master antioxidant transcription factor, in response to reflux conditions. RESULTS: We found that APE1 protein was critical for protecting against cellular ROS levels, oxidative DNA damage, double strand DNA breaks, and cell death in response to conditions that mimic reflux. Analysis of cell lines and de-identified tissues from patients with EAC demonstrated overexpression of both APE1 and NRF2 in EAC cells, as compared to non-neoplastic esophageal cells. Using reflux conditions, we detected concordant and prolonged increases of APE1 and NRF2 protein levels for several hours, following transient short exposure to ABS (20 min). NRF2 transcription activity, as measured by ARE luciferase reporter, and expression of its target genes (HO-1 and TRXND1) were similarly increased in response to ABS. Using genetic knockdown of APE1, we found that APE1 was required for the increase in NRF2 protein stability, nuclear localization, and transcription activation in EAC. Using knockdown of APE1 with reconstitution of wild-type and a redox-deficient mutant (C65A) of APE1, as well as pharmacologic APE1 redox inhibitor (E3330), we demonstrated that APE1 regulated NRF2 in a redox-dependent manner. Mechanistically, we found that APE1 is required for phosphorylation and inactivation of GSK-3beta, an important player in the NRF2 degradation pathway. CONCLUSION: APE1 redox function was required for ABS-induced activation of NRF2 by regulating phosphorylation and inactivation of GSK-3beta. The APE1-NRF2 network played a critical role in protecting esophageal cells against ROS and promoting cell survival under oxidative reflux conditions. CI - Copyright (c) 2021 The Author(s). Published by Elsevier B.V. All rights reserved. FAU - Sriramajayam, Kannappan AU - Sriramajayam K AD - Department of Surgery, Miller School of Medicine, University of Miami, Miami, FL, 33136, USA. FAU - Peng, Dunfa AU - Peng D AD - Department of Surgery, Miller School of Medicine, University of Miami, Miami, FL, 33136, USA; Sylvester Comprehensive Cancer Center, Miller School of Medicine, University of Miami, Miami, FL-33136, USA. FAU - Lu, Heng AU - Lu H AD - Department of Surgery, Miller School of Medicine, University of Miami, Miami, FL, 33136, USA. FAU - Zhou, Shoumin AU - Zhou S AD - Department of Surgery, Miller School of Medicine, University of Miami, Miami, FL, 33136, USA. FAU - Bhat, Nadeem AU - Bhat N AD - Department of Surgery, Miller School of Medicine, University of Miami, Miami, FL, 33136, USA. FAU - McDonald, Oliver G AU - McDonald OG AD - Department of Pathology, Miller School of Medicine, University of Miami, Miami, FL, 33136, USA. FAU - Que, Jianwen AU - Que J AD - Department of Medicine, Columbia University, New York, NY, 10027, USA. FAU - Zaika, Alexander AU - Zaika A AD - Department of Surgery, Miller School of Medicine, University of Miami, Miami, FL, 33136, USA; Sylvester Comprehensive Cancer Center, Miller School of Medicine, University of Miami, Miami, FL-33136, USA; Department of Veterans Affairs, Miami Healthcare System, Miami, FL, 33136, USA. FAU - El-Rifai, Wael AU - El-Rifai W AD - Department of Surgery, Miller School of Medicine, University of Miami, Miami, FL, 33136, USA; Sylvester Comprehensive Cancer Center, Miller School of Medicine, University of Miami, Miami, FL-33136, USA; Department of Veterans Affairs, Miami Healthcare System, Miami, FL, 33136, USA. Electronic address: welrifai@med.miami.edu. LA - eng GR - R01 CA138833/CA/NCI NIH HHS/United States GR - R01 CA206564/CA/NCI NIH HHS/United States GR - R01 CA224366/CA/NCI NIH HHS/United States GR - P30 CA240139/CA/NCI NIH HHS/United States GR - I01 BX002115/BX/BLRD VA/United States GR - R01 CA206563/CA/NCI NIH HHS/United States GR - IK6 BX003787/BX/BLRD VA/United States GR - I01 BX001179/BX/BLRD VA/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, U.S. Gov't, Non-P.H.S. DEP - 20210419 PL - Netherlands TA - Redox Biol JT - Redox biology JID - 101605639 RN - 0 (NF-E2-Related Factor 2) RN - 0 (NFE2L2 protein, human) RN - EC 2.7.11.1 (Glycogen Synthase Kinase 3 beta) RN - EC 4.2.99.18 (APEX1 protein, human) RN - EC 4.2.99.18 (DNA-(Apurinic or Apyrimidinic Site) Lyase) SB - IM MH - *Adenocarcinoma MH - DNA-(Apurinic or Apyrimidinic Site) Lyase MH - *Esophageal Neoplasms MH - Glycogen Synthase Kinase 3 beta MH - Humans MH - NF-E2-Related Factor 2/metabolism MH - Oxidation-Reduction PMC - PMC8082268 OTO - NOTNLM OT - APE1 OT - Acidic bile salt OT - GERD OT - GSK-3beta OT - NRF2 OT - Oxidative stress OT - ROS COIS- The authors declare no conflict of interest. EDAT- 2021/04/23 06:00 MHDA- 2021/07/06 06:00 PMCR- 2021/04/19 CRDT- 2021/04/22 20:23 PHST- 2021/01/22 00:00 [received] PHST- 2021/04/01 00:00 [revised] PHST- 2021/04/07 00:00 [accepted] PHST- 2021/04/23 06:00 [pubmed] PHST- 2021/07/06 06:00 [medline] PHST- 2021/04/22 20:23 [entrez] PHST- 2021/04/19 00:00 [pmc-release] AID - S2213-2317(21)00118-X [pii] AID - 101970 [pii] AID - 10.1016/j.redox.2021.101970 [doi] PST - ppublish SO - Redox Biol. 2021 Jul;43:101970. doi: 10.1016/j.redox.2021.101970. Epub 2021 Apr 19.