PDF(Pubmed)
Abstract：
Epithelial-mesenchymal plasticity (EMP), the process through which epithelial cells acquire mesenchymal features, is needed for wound repair but also might contribute to cancer initiation. Earlier, in vitro studies showed that Barrett\'s cells exposed to acidic bile salt solutions (ABS) develop EMP. Now, we have (1) induced reflux oesophagitis in Barrett\'s oesophagus (BO) patients by stopping proton pump inhibitors (PPIs), (2) assessed their biopsies for EMP and (3) explored molecular pathways underlying reflux-induced EMP in BO cells and spheroids.
15 BO patients had endoscopy with biopsies of Barrett\'s metaplasia while on PPIs, and 1 and 2 weeks after stopping PPIs; RNA-seq data were assessed for enrichments in hypoxia-inducible factors (HIFs), angiogenesis and EMP pathways. In BO biopsies, cell lines and spheroids, EMP features (motility) and markers (vascular endothelial growth factor (VEGF), ZEB1, miR-200a&b) were evaluated by morphology, migration assays, immunostaining and qPCR; HIF-1α was knocked down with siRNA or shRNA.
At 1 and/or 2 weeks off PPIs, BO biopsies exhibited EMP features and markers, with significant enrichment for HIF-1α, angiogenesis and EMP pathways. In BO cells, ABS induced HIF-1α activation, which decreased miR-200a&b while increasing VEGF, ZEB1 and motility; HIF-1α knockdown blocked these effects. After ABS treatment, BO spheroids exhibited migratory protrusions showing nuclear HIF-1α, increased VEGF and decreased miR-200a&b.
In BO patients, reflux oesophagitis induces EMP changes associated with increased HIF-1α signalling in Barrett\'s metaplasia. In Barrett\'s cells, ABS trigger EMP via HIF-1α signalling. Thus, HIF-1α appears to play a key role in mediating reflux-induced EMP that might contribute to cancer in BO.
NCT02579460.
15 BO patients had endoscopy with biopsies of Barrett\'s metaplasia while on PPIs, and 1 and 2 weeks after stopping PPIs; RNA-seq data were assessed for enrichments in hypoxia-inducible factors (HIFs), angiogenesis and EMP pathways. In BO biopsies, cell lines and spheroids, EMP features (motility) and markers (vascular endothelial growth factor (VEGF), ZEB1, miR-200a&b) were evaluated by morphology, migration assays, immunostaining and qPCR; HIF-1α was knocked down with siRNA or shRNA.
At 1 and/or 2 weeks off PPIs, BO biopsies exhibited EMP features and markers, with significant enrichment for HIF-1α, angiogenesis and EMP pathways. In BO cells, ABS induced HIF-1α activation, which decreased miR-200a&b while increasing VEGF, ZEB1 and motility; HIF-1α knockdown blocked these effects. After ABS treatment, BO spheroids exhibited migratory protrusions showing nuclear HIF-1α, increased VEGF and decreased miR-200a&b.
In BO patients, reflux oesophagitis induces EMP changes associated with increased HIF-1α signalling in Barrett\'s metaplasia. In Barrett\'s cells, ABS trigger EMP via HIF-1α signalling. Thus, HIF-1α appears to play a key role in mediating reflux-induced EMP that might contribute to cancer in BO.
NCT02579460.
摘要：
背景：上皮-间充质可塑性（EMP），上皮细胞获得间充质特征的过程，是伤口修复所需要的，但也可能导致癌症的发生。早些时候，体外研究表明，暴露于酸性胆汁盐溶液（ABS）的Barrett细胞会产生EMP。现在,我们有（1）通过停用质子泵抑制剂（PPI）在Barrett食管（BO）患者中诱导反流性食管炎，（2）评估其活检的EMP，（3）探索BO细胞和球体中反流诱导的EMP的分子途径。
方法：15例BO患者在使用PPI时进行了Barrett化生活检的内窥镜检查，停止PPIs后1周和2周；评估了RNA-seq数据中缺氧诱导因子（HIFs）的富集，血管生成和EMP途径。在BO活检中，细胞系和球体，EMP特征（运动性）和标志物（血管内皮生长因子（VEGF），ZEB1，miR-200a&b)通过形态学评估，迁移测定，免疫染色和qPCR;用siRNA或shRNA敲低HIF-1α。
结果：在停止PPI1周和/或2周时，BO活检显示EMP特征和标记,HIF-1α显著富集，血管生成和EMP途径。在BO细胞中,ABS诱导HIF-1α激活，它降低了miR-200a和b，同时增加了VEGF，ZEB1和运动性；HIF-1α敲低阻断了这些作用。ABS处理后，BO球体表现出迁移突起，显示核HIF-1α，VEGF增加，miR-200a和b减少。
结论：在BO患者中，反流性食管炎在Barrett's化生中诱导与HIF-1α信号增加相关的EMP变化。在巴雷特的牢房里,ABS通过HIF-1α信号触发EMP。因此，HIF-1α似乎在介导可能导致BO癌症的反流诱导的EMP中起关键作用。
背景：NCT02579460。
方法：15例BO患者在使用PPI时进行了Barrett化生活检的内窥镜检查，停止PPIs后1周和2周；评估了RNA-seq数据中缺氧诱导因子（HIFs）的富集，血管生成和EMP途径。在BO活检中，细胞系和球体，EMP特征（运动性）和标志物（血管内皮生长因子（VEGF），ZEB1，miR-200a&b)通过形态学评估，迁移测定，免疫染色和qPCR;用siRNA或shRNA敲低HIF-1α。
结果：在停止PPI1周和/或2周时，BO活检显示EMP特征和标记,HIF-1α显著富集，血管生成和EMP途径。在BO细胞中,ABS诱导HIF-1α激活，它降低了miR-200a和b，同时增加了VEGF，ZEB1和运动性；HIF-1α敲低阻断了这些作用。ABS处理后，BO球体表现出迁移突起，显示核HIF-1α，VEGF增加，miR-200a和b减少。
结论：在BO患者中，反流性食管炎在Barrett's化生中诱导与HIF-1α信号增加相关的EMP变化。在巴雷特的牢房里,ABS通过HIF-1α信号触发EMP。因此，HIF-1α似乎在介导可能导致BO癌症的反流诱导的EMP中起关键作用。
背景：NCT02579460。
