背景:胃肠上皮化生(GIM)是一种重要的癌前病变。尽管GIM的逆转具有挑战性,它可能为胃癌治疗(GC)带来一种先进的策略。缺乏合适的体外模型限制了GIM发病机制的研究,这是这项工作旨在解决进一步研究的问题。
方法:本工作采用气液界面(ALI)模型进行GIM细胞的长期培养。这项研究进行了免疫荧光(IF),定量实时聚合酶链反应(qRT-PCR),转录组测序,和粘膜蛋白质组测序(MS)技术,以确定不同群体之间差异表达基因(DEG)富集的途径,此外,验证GIM细胞的新型生物标志物。
结果:我们的研究表明GIM-ALI模型是先天GIM细胞的类似物,因此可用于粘液收集和药物筛选。我们发现基因MUC17,CDA,TRIM15、TBX3、FLVCR2、ONECUT2、ACY3、NMUR2和MAL2在GIM细胞中高表达,而GLDN,SLC5A5,MAL,和MALAT1显示下调,可用作GIM细胞的潜在生物标志物。并行,这些在GIM样本中高表达的基因主要参与癌症相关通路,如MAPK信号通路和氧化磷酸化信号通路。
结论:ALI模型首次用于GIM的体外研究。GIM-ALI模型是一种新型的体外模型,可以模拟GIM患者的组织微环境,进一步为研究GIM粘液的特征提供了途径。我们的研究确定了GIM的新标记以及与GIM相关的途径,这为探索GIM发病机制和其他潜在相关疾病提供了卓越的见解。
BACKGROUND: Gastric intestinal metaplasia (GIM) is an essential precancerous lesion. Although the reversal of GIM is challenging, it potentially brings a state-to-art strategy for gastric cancer therapeutics (GC). The lack of the appropriate in vitro model limits studies of GIM pathogenesis, which is the issue this work aims to address for further studies.
METHODS: The air-liquid interface (ALI) model was adopted for the long-term culture of GIM cells in the present work. This study conducted Immunofluorescence (IF), quantitative real-time polymerase chain reaction (qRT-PCR), transcriptomic sequencing, and mucoproteomic sequencing (MS) techniques to identify the pathways for differential expressed genes (DEGs) enrichment among different groups, furthermore, to verify novel biomarkers of GIM cells.
RESULTS: Our study suggests that GIM-ALI model is analog to the innate GIM cells, which thus can be used for mucus collection and drug screening. We found genes MUC17, CDA, TRIM15, TBX3, FLVCR2, ONECUT2, ACY3, NMUR2, and MAL2 were highly expressed in GIM cells, while GLDN, SLC5A5, MAL, and MALAT1 showed down-regulated, which can be used as potential biomarkers for GIM cells. In parallel, these genes that highly expressed in GIM samples were mainly involved in cancer-related pathways, such as the MAPK signal pathway and oxidative phosphorylation signal pathway.
CONCLUSIONS: The ALI model is validated for the first time for the in vitro study of GIM. GIM-ALI model is a novel in vitro model that can mimic the tissue micro-environment in GIM patients and further provide an avenue for studying the characteristics of GIM mucus. Our study identified new markers of GIM as well as pathways associated with GIM, which provides outstanding insight for exploring GIM pathogenesis and potentially other related conditions.