APE1 redox function is required for activation of Yes-associated protein 1 under reflux conditions in Barrett's-associated esophageal adenocarcinomas.
APE1 redox function is required for activation of Yes-associated protein 1 under reflux conditions in Barrett's-associated esophageal adenocarcinomas.
复制标题
DOI：
10.1186/s13046-022-02472-5
复制
发表时间：
2022-09-01
影响因子：
11.3
通讯作者：
El-Rifai, Wael
中科院分区：
文献类型：
--
作者：
Ballout, Farah;Lu, Heng;Chen, Lei;Sriramajayam, Kannappan;Que, Jianwen;Meng, Zhipeng;Wang, Timothy C.;Giordano, Silvia;Zaika, Alexander;McDonald, Oliver;Peng, Dunfa;El-Rifai, Wael
食管腺癌（EAC）具有预后差、生存率低的特点。慢性胃食管反流病（GERD）是巴雷特食管（BE）的主要危险因素，是一种肿瘤前化生疾病，并发展为EAC。yes相关蛋白1 （YAP1）激活介导细胞应激下的茎样特性。酸性胆汁盐（ABS）在反流条件下促进YAP1激活的作用尚未研究。本研究采用EAC细胞系、转基因小鼠和患者来源的异种移植物的组合。采用qRT-PCR、western blot、免疫组化检测APE1、YAP1 mRNA表达及蛋白水平。免疫荧光染色和荧光素酶转录活性报告试验证实YAP1激活。通过球形成实验、siRNA介导的敲低、氧化还原特异性抑制和共免疫沉淀实验确定APE1调控YAP1的功能作用和机制。我们发现，在暴露于ABS后，YAP1信号在BE和EAC细胞中被激活，ABS模拟了反流条件。这种诱导与ABS对APE1的上调一致。YAP1的激活通过其核积累和相应的YAP1靶基因的上调得到证实。ABS处理后，APE1沉默抑制了YAP1蛋白的诱导，降低了其核表达和转录活性。进一步研究发现，APE1氧化还原特异性抑制（E3330）或APE1氧化还原缺陷突变体（C65A）消除了abs介导的YAP1激活，表明APE1氧化还原依赖机制。APE1沉默或E3330处理降低了YAP1蛋白水平，减少了EAC球体的数量和大小。在机制上，我们证明了APE1通过与β-TrCP泛素酶的相互作用调节YAP1的稳定性，而APE1-氧化还原特异性抑制诱导YAP1多泛素化，促进其降解。我们的研究结果建立了APE1在EAC进展中的新功能，阐明了通过靶向APE1或YAP1治疗EAC的可药物分子脆弱性。在线版本包含补充材料，可在10.1186/s13046-022-02472-5获得。
Esophageal adenocarcinoma (EAC) is characterized by poor prognosis and low survival rate. Chronic gastroesophageal reflux disease (GERD) is the main risk factor for the development of Barrett’s esophagus (BE), a preneoplastic metaplastic condition, and its progression to EAC. Yes-associated protein 1 (YAP1) activation mediates stem-like properties under cellular stress. The role of acidic bile salts (ABS) in promoting YAP1 activation under reflux conditions remains unexplored. A combination of EAC cell lines, transgenic mice, and patient-derived xenografts were utilized in this study. mRNA expression and protein levels of APE1 and YAP1 were evaluated by qRT-PCR, western blot, and immunohistochemistry. YAP1 activation was confirmed by immunofluorescence staining and luciferase transcriptional activity reporter assay. The functional role and mechanism of regulation of YAP1 by APE1 was determined by sphere formation assay, siRNA mediated knockdown, redox-specific inhibition, and co-immunoprecipitation assays. We showed that YAP1 signaling is activated in BE and EAC cells following exposure to ABS, the mimicry of reflux conditions in patients with GERD. This induction was consistent with APE1 upregulation in response to ABS. YAP1 activation was confirmed by its nuclear accumulation with corresponding up-regulation of YAP1 target genes. APE1 silencing inhibited YAP1 protein induction and reduced its nuclear expression and transcriptional activity, following ABS treatment. Further investigation revealed that APE1-redox-specific inhibition (E3330) or APE1 redox-deficient mutant (C65A) abrogated ABS-mediated YAP1 activation, indicating an APE1 redox-dependent mechanism. APE1 silencing or E3330 treatment reduced YAP1 protein levels and diminished the number and size of EAC spheroids. Mechanistically, we demonstrated that APE1 regulated YAP1 stability through interaction with β-TrCP ubiquitinase, whereas APE1-redox-specific inhibition induced YAP1 poly-ubiquitination promoting its degradation. Our findings established a novel function of APE1 in EAC progression elucidating druggable molecular vulnerabilities via targeting APE1 or YAP1 for the treatment of EAC. The online version contains supplementary material available at 10.1186/s13046-022-02472-5.
登录
查看更多内容
影响因子：
50.3
作者：
Quante M;Bhagat G;Abrams JA;Marache F;Good P;Lee MD;Lee Y;Friedman R;Asfaha S;Dubeykovskaya Z;Mahmood U;Figueiredo JL;Kitajewski J;Shawber C;Lightdale CJ;Rustgi AK;Wang TC
通讯作者：
Wang TC
影响因子：
11.2
作者：
Jorgenson TC;Zhong W;Oberley TD
通讯作者：
Oberley TD
影响因子：
4.7
作者：
Bahmad HF;Cheaito K;Chalhoub RM;Hadadeh O;Monzer A;Ballout F;El-Hajj A;Mukherji D;Liu YN;Daoud G;Abou-Kheir W
通讯作者：
Abou-Kheir W
DOI：
10.1158/1078-0432.ccr-14-2191
发表时间：
2015-06-01
期刊：
Clinical cancer research : an official journal of the American Association for Cancer Research
影响因子：
--
作者：
Song S;Honjo S;Jin J;Chang SS;Scott AW;Chen Q;Kalhor N;Correa AM;Hofstetter WL;Albarracin CT;Wu TT;Johnson RL;Hung MC;Ajani JA
通讯作者：
Ajani JA
影响因子：
11.2
作者：
Lu, Heng;Bhat, Ajaz A.;El-Rifai, Wael
通讯作者：
El-Rifai, Wael