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.
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。