Wang Hailei, Lou Chunyan, Ma Na
Department of Pediatrics, Huaihe Hospital of Henan University, Kaifeng, Henan, China.
Department of Neurology, Henan Kaifeng Children's Hospital, Kaifeng, Henan, China,
Cancer Manag Res. 2019 Feb 19;11:1641-1651. doi: 10.2147/CMAR.S177508. eCollection 2019.
Although many miRNAs are identified to be deregulated and play vital roles in the progression of Wilms' tumor (WT), there are still a large number of miRNAs are waiting for us to explore. The purpose of the present study is to investigate the different expressing profiles of miRNAs in WT tissues and the adjacent normal tissues, and probe the effects and mechanism of a certain miRNA among the different expressing miRNAs.
miRNA microarray was recruited to assess the differently expressed miRNAs in WT tissues and normal tissues, which was further verified by RT-PCR. Receiver operating characteristic curves were performed to calculate the specificity and sensitivity of miRNAs in the diagnose of WT. CCK-8, flow cytometry, wound healing, transwell chamber and tumor-burdened assays were used to assess cell growth, apoptosis, migration, invasion and tumorigenesis. Luciferase report assay was used to evaluate the interaction between miR-140-5p and .
A total of 34 miRNAs were abnormally expressed in the WT tissues, among which, miR-140-5p was identified to be obviously down-regulated in WT tissues, and the AUC of it was 0.961. Besides, we found that patients with miR-140-5p low expression always had a shorter overall survival and more aggressive clinical features, such as bigger tumor size (=0.002), higher pathological stage (=0.003) and higher occurrence rate of lymph node metastasis (=0.009) than those in patients with miR-140-5p high expression. Moreover, luciferase reporter assay showed that was the direct target of miR-140-5p, which was negatively regulated by miR-140-5p and was highly expressed in WT tissues. Furthermore, knockdown of miR-140-5p obviously enhanced the proliferation and tumorigenesis and repressed the apoptosis of G401 cells, and these effects were all abolished when was down-regulated.
The present study illustrates that miR-140-5p functions as a tumor suppressor in the occurrence and development of WT via targeting , which provides theoretical foundation for serving miR-140-5p as a new diagnosis marker even a therapeutic target for WT.
尽管已鉴定出许多微小RNA（miRNA）在肾母细胞瘤（WT）进展过程中失调并发挥重要作用，但仍有大量miRNA有待探索。本研究旨在调查WT组织和相邻正常组织中miRNA的不同表达谱，并探究不同表达的miRNA中某一特定miRNA的作用及机制。
采用miRNA芯片评估WT组织和正常组织中差异表达的miRNA，并通过逆转录-聚合酶链反应（RT-PCR）进一步验证。绘制受试者工作特征曲线以计算miRNA在WT诊断中的特异性和敏感性。采用细胞计数试剂盒-8（CCK-8）、流式细胞术、伤口愈合实验、Transwell小室实验和荷瘤实验评估细胞生长、凋亡、迁移、侵袭和肿瘤发生情况。采用荧光素酶报告基因实验评估miR-140-5p与……之间的相互作用。
WT组织中共有34种miRNA异常表达，其中miR-140-5p在WT组织中明显下调，其曲线下面积（AUC）为0.961。此外，我们发现miR-140-5p低表达的患者总生存期较短，且具有更具侵袭性的临床特征，如肿瘤体积更大（P = 0.002）、病理分期更高（P = 0.003）和淋巴结转移发生率更高（P = 0.009），相比于miR-140-5p高表达的患者。此外，荧光素酶报告基因实验表明……是miR-140-5p的直接靶点，其受到miR-140-5p的负调控且在WT组织中高表达。此外，敲低miR-140-5p明显增强了G401细胞的增殖和肿瘤发生能力，并抑制了其凋亡，而当……下调时，这些作用均被消除。
本研究表明miR-140-5p通过靶向……在WT的发生发展中发挥肿瘤抑制作用，这为将miR-140-5p作为WT的新诊断标志物甚至治疗靶点提供了理论基础。
