Department of Molecular Medicine, Université Laval, Québec, QC PQ G1V 0A6, Canada.
Research Center of Quebec CHU-Université Laval, Québec, QC PQ G1E6W2, Canada.
Int J Mol Sci. 2020 Mar 7;21(5):1848. doi: 10.3390/ijms21051848.
Growing evidence demonstrates that epithelial-mesenchymal transition (EMT) plays an important role in epithelial ovarian cancer (EOC) progression and spreading; however, its molecular mechanisms remain poorly defined. We have previously shown that the antigen receptor LY75 can modulate EOC cell phenotype and metastatic potential, as LY75 depletion directed mesenchymal-epithelial transition (MET) in EOC cell lines with mesenchymal phenotype. We used the LY75-mediated modulation of EMT as a model to investigate for DNA methylation changes during EMT in EOC cells, by applying the reduced representation bisulfite sequencing (RRBS) methodology. Numerous genes have displayed EMT-related DNA methylation patterns alterations in their promoter/exon regions. Ten selected genes, whose DNA methylation alterations were further confirmed by alternative methods, were further identified, some of which could represent new EOC biomarkers/therapeutic targets. Moreover, our methylation data were strongly indicative for the predominant implication of the Wnt/β-catenin pathway in the EMT-induced DNA methylation variations in EOC cells. Consecutive experiments, including alterations in the Wnt/β-catenin pathway activity in EOC cells with a specific inhibitor and the identification of LY75-interacting partners by a proteomic approach, were strongly indicative for the direct implication of the LY75 receptor in modulating the Wnt/β-catenin signaling in EOC cells.
越来越多的证据表明上皮-间充质转化（EMT）在卵巢上皮癌（EOC）的进展和扩散中起着重要作用；然而，其分子机制仍未明确定义。我们之前已经表明，抗原受体 LY75 可以调节 EOC 细胞表型和转移潜能，因为 LY75 耗竭可指导具有间充质表型的 EOC 细胞系发生间充质-上皮转化（MET）。我们使用 LY75 介导的 EMT 调节作为模型，通过应用简化代表性亚硫酸氢盐测序（RRBS）方法，研究 EMT 过程中 EOC 细胞中的 DNA 甲基化变化。许多基因在其启动子/外显子区域显示出与 EMT 相关的 DNA 甲基化模式改变。选择了十个 DNA 甲基化改变进一步通过其他方法确认的基因，其中一些可能代表新的 EOC 生物标志物/治疗靶点。此外，我们的甲基化数据强烈表明 Wnt/β-catenin 通路在 EMT 诱导的 EOC 细胞 DNA 甲基化变化中占主导地位。后续实验包括在 EOC 细胞中改变 Wnt/β-catenin 通路活性的实验以及通过蛋白质组学方法鉴定 LY75 相互作用伙伴的实验，都强烈表明 LY75 受体直接参与调节 EOC 细胞中的 Wnt/β-catenin 信号通路。
