Hong Liyi, Zhao Xu, Shao Xuejun, Zhu Hong
Clinical Medical Laboratory, Children's Hospital of Soochow University, Suzhou, Jiangsu 215004, P.R. China.
Mol Med Rep. 2017 Jul;16(1):247-253. doi: 10.3892/mmr.2017.6561. Epub 2017 May 10.
Nephroblastoma (Wilms' tumor) is frequently associated with mortality in children. MicroRNAs (miRNAs) are important for tumor development serving as oncogenes or tumor suppressors. In the present study, miRNA‑590 (miR‑590) was identified to be upregulated in Wilms' tumor tissues compared with the normal adjacent tissues. Additionally, the levels of miR‑590 were consistent with their clinical stage. Wilms' tumor 1 (WT1) was considered to be a tumor suppressor in certain tumor types, and it has been detected at low expression levels in various types of cancer with high cell proliferation and aggressive behavior. The expression levels of miR‑590 were quantified using reverse transcription‑quantitative polymerase chain reaction. Cell proliferation was measured using 5‑ethynyl‑20‑deoxyuridine assays. The protein expression levels of WT1 were investigated by western blot analysis. To the best of our knowledge, the present study was the first to determine that WT1 was a target gene of miR‑590 as miR‑590 was able to negatively regulate WT1 expression level by binding to the specific target site within the 3'‑untranslated region (3'‑UTR) of WT1 in G401 cells. Additionally, overexpression of miR‑590 promoted G401 cell proliferation which was consistent with the effect of small interfering RNA‑WT1. Subsequently, the present study determined that the cell phenotype altered by miR‑590 overexpression may be reversed by upregulation of WT1 in G401 cells. In conclusion, the observations indicated that miR‑590 may function as an oncogene via targeting WT1 to induce G401 cell proliferation. These results may contribute to current understanding of the function of miR‑590 in nephroblastoma.
肾母细胞瘤(威尔姆斯瘤)常导致儿童死亡。微小RNA(miRNA)作为癌基因或肿瘤抑制因子,对肿瘤发展至关重要。在本研究中,与相邻正常组织相比,miRNA-590(miR-590)在威尔姆斯瘤组织中被鉴定为上调。此外,miR-590的水平与其临床分期一致。威尔姆斯瘤1(WT1)在某些肿瘤类型中被认为是一种肿瘤抑制因子,并且在各种具有高细胞增殖和侵袭性行为的癌症中检测到其低表达水平。使用逆转录-定量聚合酶链反应对miR-590的表达水平进行定量。使用5-乙炔基-2'-脱氧尿苷检测法测量细胞增殖。通过蛋白质印迹分析研究WT1的蛋白质表达水平。据我们所知,本研究首次确定WT1是miR-590的靶基因,因为miR-590能够通过与G401细胞中WT1的3'-非翻译区(3'-UTR)内的特定靶位点结合来负调节WT1的表达水平。此外,miR-590的过表达促进了G401细胞增殖,这与小干扰RNA-WT1的作用一致。随后,本研究确定在G401细胞中上调WT1可逆转miR-590过表达所改变的细胞表型。总之,这些观察结果表明miR-590可能通过靶向WT1诱导G401细胞增殖而发挥癌基因的作用。这些结果可能有助于当前对miR-590在肾母细胞瘤中功能的理解。