Table1_Evaluation of the lncRNA-miRNA-mRNA ceRNA network in lungs of miR-147 −/− mice.docx

Background: Previous studies have documented important roles for microRNA-147 (miR-147) in inflammation, radiation-induced injury, cancer, and a range of other diseases. Murine lungs exhibit high levels of miRNA, mRNA, and lncRNA expression. However, very little research to date has focused on the lncRNA-miRNA-mRNA competing endogenous RNA (ceRNA) networks associated with miR-147, and the regulation of lncRNAs and miRNAs in this setting remains poorly understood.
Methods: After establishing a miR-147−/− model mouse, samples of lung tissue were harvested for RNA-sequencing, and differentially expressed lncRNAs, miRNAs, and mRNAs were identified. The miRNA targets of these lncRNAs and the identified miRNAs were first overlapped to facilitate the prediction of target mRNAs, with analyses then examining the overlap between these targets and mRNAs that were differentially expressed. Then, these target mRNAs were subjected to pathway enrichment analyses. These results were ultimately used to establish a miR-147-related ceRNA network.
Results: Relative to wild-type mice, the lungs of miR-147−/− mice exhibited 91, 43, and 71 significantly upregulated lncRNAs, miRNAs, and mRNAs, respectively, together with 114, 31, and 156 that were significantly downregulated. The lncRNA-miRNA-mRNA network established based on these results led to the identification of Kcnh6 as a differentially expressed hub gene candidate and enabled the identification of a range of regulatory relationships. KEGG pathway enrichment showed that the mRNA targets of differentially expressed lncRNAs and miRNAs in the mice were associated with tumor-related signaling, endometrial cancer, bladder cancer, and ErbB signaling.
Conclusion: These results suggest that the identified ceRNA network in miR-147−/− mice shapes tumor-associated signaling activity, with miR-147 potentially regulating various lncRNAs and miRNAs through Kcnh6, ultimately influencing tumorigenesis. Future studies of the lncRNA, miRNA, and mRNA regulatory targets shown to be associated with miR-147 in the present study may ultimately lead to the identification of novel clinically relevant targets through which miR-147 shapes the pathogenesis of cancer and other diseases.

背景：既往研究已证实微小RNA-147（microRNA-147，miR-147）在炎症、辐射诱导损伤、癌症及多种其他疾病中发挥重要作用。小鼠肺部呈现高水平的微小RNA（microRNA）、信使RNA（mRNA）及长链非编码RNA（lncRNA）表达。然而，迄今为止鲜有研究聚焦于与miR-147相关的长链非编码RNA-微小RNA-信使RNA竞争性内源RNA（ceRNA）调控网络，该场景下长链非编码RNA与微小RNA的调控机制仍有待阐明。
方法：本研究构建miR-147基因敲除（miR-147−/−）小鼠模型，采集肺组织样本进行RNA测序（RNA-sequencing），鉴定差异表达的长链非编码RNA、微小RNA及信使RNA。首先通过重叠分析上述长链非编码RNA与微小RNA的靶标微小RNA及靶标信使RNA，以预测靶标信使RNA；随后进一步分析这些靶标与差异表达信使RNA的重叠部分。随后对所得靶标信使RNA进行通路富集分析。最终基于上述结果构建与miR-147相关的竞争性内源RNA调控网络。
结果：与野生型小鼠相比，miR-147基因敲除小鼠的肺组织中分别有91个、43个及71个长链非编码RNA、微小RNA及信使RNA显著上调，另有114个、31个及156个显著下调。基于上述结果构建的长链非编码RNA-微小RNA-信使RNA调控网络，鉴定出Kcnh6作为差异表达核心枢纽基因候选者，并明确了一系列调控关系。京都基因与基因组百科全书（KEGG）通路富集分析显示，该基因敲除小鼠中差异表达的长链非编码RNA与微小RNA的靶标信使RNA与肿瘤相关信号通路、子宫内膜癌、膀胱癌及ErbB信号通路（ErbB signaling）密切相关。
结论：本研究结果表明，miR-147基因敲除小鼠中鉴定出的竞争性内源RNA调控网络可调控肿瘤相关信号通路活性，miR-147或可通过Kcnh6调控多种长链非编码RNA与微小RNA的表达，最终影响肿瘤发生进程。未来针对本研究中与miR-147相关的长链非编码RNA、微小RNA及信使RNA调控靶点开展研究，或可鉴定出全新的临床相关靶点，进而阐明miR-147在癌症及其他疾病发病机制中的作用。