Abstract：
BACKGROUND: The pathogenesis of biliary atresia (BA) remains elusive. We aimed to investigate the role of long noncoding RNA (lncRNA) MEG9 in BA.
METHODS: LncRNA microarray was conducted to identify differentially expressed lncRNAs in three BA and three para-hepatoblastoma liver tissues. RT-qPCR validated the results. Human intrahepatic bile duct epithelial cells (HIBECs) were stably transfected with lncRNA MEG9 knockdown/overexpression to investigate its cellular localization and function. RNA sequencing (RNA-seq), differentially expressed genes (DEGs) analysis and gene set enrichment analysis were applied to MEG9-overexpresed HIBECs. RNA pull-down and mass spectrometry explored the interacting protein of MEG9, while clinical information was reviewed.
RESULTS: 436 differentially expressed lncRNAs were identified, with MEG9 highly upregulated in BA. RT-qPCR further confirmed MEG9\'s overexpression in BA and diagnostic potential (AUC = 0.9691). MEG9 was predominantly located in the nucleus and significantly promoted cell proliferation and migration. RNA-seq revealed inflammation- and extracellular matrix-related pathways enriched in MEG9-overexpressing HIBECs, with upregulated cytokine genes like CXCL6 and IL6. MMP-7 and collagen I were also overexpressed. Furthermore, 38 proteins were identified to specifically interact with MEG9, and S100A9 was highly expressed in cell models. S100A9 was also significantly upregulated in BA liver tissue and correlated with MEG9 expression (r = 0.313, p < 0.05), albumin level (r = -0.349, p < 0.05), and platelet level (r = -0.324, p < 0.05).
CONCLUSIONS: MEG9 influences cholangiocyte proliferation, migration, and cytokine production, potentially regulating BA inflammation and fibrosis via S100A9 interaction.
METHODS: LncRNA microarray was conducted to identify differentially expressed lncRNAs in three BA and three para-hepatoblastoma liver tissues. RT-qPCR validated the results. Human intrahepatic bile duct epithelial cells (HIBECs) were stably transfected with lncRNA MEG9 knockdown/overexpression to investigate its cellular localization and function. RNA sequencing (RNA-seq), differentially expressed genes (DEGs) analysis and gene set enrichment analysis were applied to MEG9-overexpresed HIBECs. RNA pull-down and mass spectrometry explored the interacting protein of MEG9, while clinical information was reviewed.
RESULTS: 436 differentially expressed lncRNAs were identified, with MEG9 highly upregulated in BA. RT-qPCR further confirmed MEG9\'s overexpression in BA and diagnostic potential (AUC = 0.9691). MEG9 was predominantly located in the nucleus and significantly promoted cell proliferation and migration. RNA-seq revealed inflammation- and extracellular matrix-related pathways enriched in MEG9-overexpressing HIBECs, with upregulated cytokine genes like CXCL6 and IL6. MMP-7 and collagen I were also overexpressed. Furthermore, 38 proteins were identified to specifically interact with MEG9, and S100A9 was highly expressed in cell models. S100A9 was also significantly upregulated in BA liver tissue and correlated with MEG9 expression (r = 0.313, p < 0.05), albumin level (r = -0.349, p < 0.05), and platelet level (r = -0.324, p < 0.05).
CONCLUSIONS: MEG9 influences cholangiocyte proliferation, migration, and cytokine production, potentially regulating BA inflammation and fibrosis via S100A9 interaction.
摘要：
背景：胆道闭锁（BA）的发病机制仍然难以捉摸。我们的目的是研究长链非编码RNA（lncRNA）MEG9在BA中的作用。
方法：进行LncRNA微阵列以鉴定三种BA和三种肝旁母细胞瘤肝组织中差异表达的lncRNA。RT-qPCR验证了结果。用lncRNAMEG9敲低/过表达稳定转染人肝内胆管上皮细胞（HIBECs），以研究其细胞定位和功能。RNA测序（RNA-seq），将差异表达基因（DEGs）分析和基因集富集分析应用于MEG9过表达的HIBECs。RNA下拉和质谱研究了MEG9的相互作用蛋白，同时回顾了临床信息。
结果：确定了436个差异表达的lncRNAs，MEG9在BA中高度上调。RT-qPCR进一步证实了BA中MEG9的过表达和诊断潜力(AUC=0.9691)。MEG9主要位于细胞核中，并显着促进细胞增殖和迁移。RNA-seq揭示了富含MEG9过表达HIBECs的炎症和细胞外基质相关途径，与细胞因子基因如CXCL6和IL6上调。MMP-7和胶原蛋白I也过表达。此外，鉴定了38种蛋白质与MEG9特异性相互作用，并且S100A9在细胞模型中高度表达。S100A9在BA肝组织中也显著上调,与MEG9表达呈正相关(r=0.313,p<0.05),白蛋白水平（r=-0.349，p<0.05），和血小板水平（r=-0.324，p<0.05）。
结论：MEG9影响胆管细胞增殖，迁移,和细胞因子的产生，可能通过S100A9相互作用调节BA炎症和纤维化。
方法：进行LncRNA微阵列以鉴定三种BA和三种肝旁母细胞瘤肝组织中差异表达的lncRNA。RT-qPCR验证了结果。用lncRNAMEG9敲低/过表达稳定转染人肝内胆管上皮细胞（HIBECs），以研究其细胞定位和功能。RNA测序（RNA-seq），将差异表达基因（DEGs）分析和基因集富集分析应用于MEG9过表达的HIBECs。RNA下拉和质谱研究了MEG9的相互作用蛋白，同时回顾了临床信息。
结果：确定了436个差异表达的lncRNAs，MEG9在BA中高度上调。RT-qPCR进一步证实了BA中MEG9的过表达和诊断潜力(AUC=0.9691)。MEG9主要位于细胞核中，并显着促进细胞增殖和迁移。RNA-seq揭示了富含MEG9过表达HIBECs的炎症和细胞外基质相关途径，与细胞因子基因如CXCL6和IL6上调。MMP-7和胶原蛋白I也过表达。此外，鉴定了38种蛋白质与MEG9特异性相互作用，并且S100A9在细胞模型中高度表达。S100A9在BA肝组织中也显著上调,与MEG9表达呈正相关(r=0.313,p<0.05),白蛋白水平（r=-0.349，p<0.05），和血小板水平（r=-0.324，p<0.05）。
结论：MEG9影响胆管细胞增殖，迁移,和细胞因子的产生，可能通过S100A9相互作用调节BA炎症和纤维化。
