Activation of NRF2 by APE1/REF1 is redox-dependent in Barrett's related esophageal adenocarcinoma cells.
Activation of NRF2 by APE1/REF1 is redox-dependent in Barrett's related esophageal adenocarcinoma cells.
复制标题
DOI：
10.1016/j.redox.2021.101970
复制
发表时间：
2021-07
期刊：
影响因子：
11.4
通讯作者：
El-Rifai W
中科院分区：
文献类型：
--
作者：
Sriramajayam K;Peng D;Lu H;Zhou S;Bhat N;McDonald OG;Que J;Zaika A;El-Rifai W
慢性胃食管反流病（GERD）是化生性Barrett食管（BE）发展及其进展为食管腺癌（EAC）的主要危险因素。在回流条件下，响应于酸性胆汁盐（ABS）的活性氧（ROS）的不受控制的积累可能对细胞是致命的。在这项研究中，我们研究了APE 1/REF 1在调节核红细胞样因子2（NRF 2），主抗氧化转录因子，在回流条件下的作用。我们发现，APE 1蛋白是至关重要的保护细胞的ROS水平，氧化DNA损伤，双链DNA断裂，细胞死亡的条件下，模拟反流。对EAC患者的细胞系和去识别组织的分析表明，与非肿瘤性食管细胞相比，EAC细胞中APE 1和NRF 2均过表达。使用回流条件下，我们检测到一致的和延长的APE 1和NRF 2蛋白水平的增加数小时，短暂的短期暴露于ABS（20分钟）。通过ARE荧光素酶报告基因测量的NRF 2转录活性及其靶基因（HO-1和TRXND 1）的表达在ABS的反应中类似地增加。利用APE 1的基因敲低，我们发现APE 1是增加EAC中NRF 2蛋白稳定性、核定位和转录激活所必需的。利用APE 1的敲除与野生型和APE 1的氧化还原缺陷突变体（C65 A）的重建，以及药理学APE 1氧化还原抑制剂（E3330），我们证明了APE 1以氧化还原依赖性方式调节NRF 2。从机制上讲，我们发现APE 1是GSK-3β磷酸化和失活所必需的，GSK-3β是NRF 2降解途径中的重要参与者。APE 1的氧化还原功能通过调节GSK-3β的磷酸化和失活而参与ABS诱导的NRF 2活化。APE 1-NRF 2网络在保护食管细胞免受ROS和促进氧化反流条件下的细胞存活方面发挥关键作用。APE 1保护食管肿瘤细胞免受酸性胆汁盐（ABS）诱导的氧化应激、DNA损伤和细胞死亡。APE 1通过介导抗氧化转录因子NRF 2的活性来调节氧化应激。APE 1主要通过GSK-3β维持NRF 2在细胞核中的稳定性并抑制NRF 2蛋白降解。APE 1的氧化还原功能是GSK-3β的适当磷酸化和失活以及随后的NRF 2活化所必需的。
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. 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-3β, an important player in the NRF2 degradation pathway. APE1 redox function was required for ABS-induced activation of NRF2 by regulating phosphorylation and inactivation of GSK-3β. The APE1-NRF2 network played a critical role in protecting esophageal cells against ROS and promoting cell survival under oxidative reflux conditions. APE1 protected esophageal neoplastic cells from acidic bile salts (ABS)-induced oxidative stress, DNA damage and cell death. APE1 regulated oxidative stress through mediating the activity of master antioxidant transcription factor, NRF2. APE1 was required to maintain NRF2 stability in the nucleus and suppressed NRF2 protein degradation, mainly through GSK-3β. APE1 redox function was required for proper phosphorylation and inactivation of GSK-3β, and subsequent activation of NRF2.
登录
查看更多内容
影响因子：
4.7
作者：
Jenkins, Gareth J. S.;D'Souza, Francis R.;Baxter, John N.
通讯作者：
Baxter, John N.
影响因子：
11.2
作者：
Lu, Heng;Bhat, Ajaz A.;El-Rifai, Wael
通讯作者：
El-Rifai, Wael
影响因子：
16
作者：
Padmanabhan, B;Tong, KI;Yamamoto, M
通讯作者：
Yamamoto, M
影响因子：
5
作者：
Le Bras, Gregoire F.;Loomans, Holli A.;Andl, Claudia D.
通讯作者：
Andl, Claudia D.
影响因子：
2.9
作者：
He, Haiqi;Chen, Nanzheng;Fu, Junke
通讯作者：
Fu, Junke