Abstract：
Barrett\'s esophagus (BE) belongs to a pathological phenomenon occurring in the esophagus, this paper intended to unveil the underlying function of miR-378a-5p and its target TSPAN8 in BE progression. GEO analysis was conducted to determine differentially expressed genes in BE samples. Non-dysplastic metaplasia BE samples, high-grade dysplastic BE samples and controls were collected from subjects. CP-A and CP-B cells were exposed to bile acids (BA) to mimic gastroesophageal reflux in BE cells. RT-qPCR as well as western blot were applied for verifying expressions of miR-378a-5p, TSPAN8, CDX2 and SOX9. CCK-8, wound scratch together with Transwell assays were exploited for ascertaining cell proliferation, migration as well as invasion. The targeted relationship of miR-378a-5p and TSPAN8 could be verified by correlation analysis, dual-luciferase reporter experiment, and rescue experiments. Through analyzing GSE26886 dataset, we screened the most abundantly expressed gene TSPAN8 in BE samples. miR-378a-5p was reduced whereas TSPAN8 was elevated in CP-A as well as CP-B cells after triggering with BA. Knocking down TSPAN8 could counteract BA-triggered enhancement in BE cell proliferation, migration along with invasion. miR-378a-5p could suppress BE cell proliferation, and migration along with invasion via targeting TSPAN8. In BE, miR-378a-5p targeted TSPAN8 to inhibit BE cell proliferation, and migration along invasion. miR-378a-5p deletion or elevation of TSPAN8 may be key point in regulating CDX2 and SOX9 levels, thereby promoting BE formation.
摘要：
Barrett食管（BE）属于发生在食管中的病理现象，本文旨在揭示miR-378a-5p及其靶标TSPAN8在BE进展中的潜在功能。进行GEO分析以确定BE样品中差异表达的基因。非发育不良化生BE样本，从受试者中收集高级别发育不良BE样品和对照。将CP-A和CP-B细胞暴露于胆汁酸(BA)以模拟BE细胞中的胃食管反流。RT-qPCR和Westernblot用于验证miR-378a-5p的表达。TSPAN8、CDX2和SOX9。CCK-8，伤口划痕与Transwell测定一起用于确定细胞增殖，移民和入侵。通过相关性分析可以验证miR-378a-5p与TSPAN8的靶向关系,双荧光素酶报告基因实验，和救援实验。通过分析GSE26886数据集，我们筛选了BE样品中表达最丰富的基因TSPAN8。在用BA触发后，miR-378a-5p降低，而TSPAN8在CP-A和CP-B细胞中升高。敲除TSPAN8可以抵消BA引发的BE细胞增殖增强，移民和入侵。miR-378a-5p可以抑制BE细胞增殖，以及通过靶向TSPAN8的迁移和入侵。在BE中,miR-378a-5p靶向TSPAN8抑制BE细胞增殖，以及沿着入侵迁移。miR-378a-5p缺失或TSPAN8升高可能是调控CDX2和SOX9水平的关键点,从而促进BE的形成。
