Zhang Hong-Mei, Cui Ming-Yu, Chen Zhi-Hong
Department of Urology, Qilu Children's Hospital of Shandong University, Jinan 250022, Shandong Province,People's Republic of China.
J Genet. 2023;102.
Wilms' tumour (WT) is the most typical type of renal tumour in children, which has a poor prognosis and high recurrence rate. This study explored whether lncRNA EMX2 opposite strand / antisense RNA (EMX2OS) modulated the stemness, epithelial-mesenchymal transition (EMT) and metastasis of WTcells through the interaction with insulin-like growth factor 2 mRNA-binding protein 1 (IGF2BP1). The expression levels of EMX2OS, IGF2BP1 and stem cell markers OCT4, Nanog, Sox2 and CD133 were detected by real time quantitative polymerase chain reaction (RT-qPCR). The stemness, migration and invasion of WTcells were determined by sphere formation assay, scratch and transwell assay, respectively. The levels of EMT-related proteins were detected by Western blotting. RNA pull down and RIP assays were utilized to validate the interaction between EMX2OS and IGF2BP1. The tumourigenicity of WTcells was analysed using a xenograft tumour assay. EMX2OS was downregulated in WT patients, while IGF2BP1 was upregulated. EMX2OS overexpression or IGF2BP1 knockdown suppressed WT cell sphere formation, migration and invasion. Moreover, EMX2OS could directly interact with RNA-binding protein IGF2BP1, and IGF2BP1 overexpression counteracted the inhibitory effect of EMX2OS on WT cell stemness, migration, invasion and EMT. The tumour growth, stemness and EMT were repressed by EMX2OS through the interaction with IGF2BP1. In conclusion, EMX2OS acted as a tumour suppressor for WT by interacting with IGF2BP1, which might be a novel target for WT diagnosis and therapy.
肾母细胞瘤（WT）是儿童最典型的肾肿瘤类型，其预后较差且复发率高。本研究探讨了长链非编码RNA EMX2反义链/反义RNA（EMX2OS）是否通过与胰岛素样生长因子2 mRNA结合蛋白1（IGF2BP1）相互作用来调节WT细胞的干性、上皮-间质转化（EMT）和转移。通过实时定量聚合酶链反应（RT-qPCR）检测EMX2OS、IGF2BP1以及干细胞标志物OCT4、Nanog、Sox2和CD133的表达水平。分别通过成球试验、划痕试验和Transwell试验测定WT细胞的干性、迁移和侵袭能力。通过蛋白质免疫印迹法检测EMT相关蛋白的水平。利用RNA下拉和RNA免疫沉淀试验验证EMX2OS与IGF2BP1之间的相互作用。使用异种移植肿瘤试验分析WT细胞的致瘤性。WT患者中EMX2OS表达下调，而IGF2BP1表达上调。EMX2OS过表达或IGF2BP1敲低可抑制WT细胞成球、迁移和侵袭。此外，EMX2OS可直接与RNA结合蛋白IGF2BP1相互作用，IGF2BP1过表达可抵消EMX2OS对WT细胞干性、迁移、侵袭和EMT的抑制作用。EMX2OS通过与IGF2BP1相互作用抑制肿瘤生长、干性和EMT。总之，EMX2OS通过与IGF2BP1相互作用作为WT的肿瘤抑制因子，这可能是WT诊断和治疗的新靶点。
