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Abstract：
Wilms tumor (WT) is the most common solid childhood tumors all over the world. MicroRNAs (miRs) contribute to tumorigenesis of various cancers through targeting gene. The present study investigated the vital role of miR-194-5p and its underlying mechanism in the progression of WT. Immunohistochemistry and quantitative real-time polymerase chain reaction (qRT-PCR) assay indicated downregulation of miR194-5p and upregulation of Crk, in WT tissues compared to adjacent normal tissues. Transfection with miR-194-5p mimics into nephroblastoma cells showed a significant decline in cell migration and invasion, which was detected by Transwell assay. Luciferase assay confirmed that Crk was a direct target gene of miR-194-5p. More important, the mesenchymal to epithelial transition (EMT) biomarkers containing E-cadherin, N-cadherin and Zeb1 were examined by Western blot, and revealed that miR-194-5p mimics decreased the levels of N-cadherin and Zeb1 but increased E-cadherin, which suggested that miR-194-5p inhibited EMT. Crk knockdown could reverse the increased nephroblastoma cell invasion, migration and EMT caused by miR-194-5p inhibitor. Interestingly, qRT-PCR and Western blot analysis showed that overexpression of miR-194-5p deactivated HGF/c-Met/Scr signaling pathway via targeting Crk. In conclusion, miR-194-5p inhibited nephroblastoma cell metastasis and EMT in the progression of WT by targeting Crk. Thus, miR-194-5p might be a potential target in WT particularly for the prevention of metastasis and EMT.
摘要：
Wilms肿瘤（WT）是世界上最常见的实体儿童肿瘤。MicroRNAs(miRs)通过靶向基因参与多种癌症的肿瘤发生。本研究探讨了miR-194-5p在WT进展中的重要作用及其潜在机制。免疫组织化学和定量实时聚合酶链反应（qRT-PCR）分析显示miR194-5p下调和Crk上调，在WT组织中与邻近正常组织相比。用miR-194-5p模拟物转染肾母细胞瘤细胞显示细胞迁移和侵袭能力显著下降,通过Transwell检测。荧光素酶检测证实Crk是miR-194-5p的直接靶基因。更重要的是,含有E-cadherin的间充质向上皮转化（EMT）生物标志物，通过蛋白质印迹检查N-钙黏着蛋白和Zeb1，并揭示miR-194-5p模拟物降低了N-cadherin和Zeb1的水平，但增加了E-cadherin，这表明miR-194-5p抑制EMT。Crk敲低可以逆转肾母细胞瘤细胞侵袭的增加，miR-194-5p抑制剂引起的迁移和EMT。有趣的是,qRT-PCR和Westernblot分析显示miR-194-5p的过表达通过靶向Crk使HGF/c-Met/Scr信号通路失活。总之,miR-194-5p通过靶向Crk抑制WT进展中的肾母细胞瘤细胞转移和EMT。因此，miR-194-5p可能是WT中的潜在靶标，特别是用于预防转移和EMT。
