Department of Biochemistry & Immunology, Capital Institute of Pediatrics, NO. 2, Yabao Road, Chaoyang District, Beijing, 100020, China.
Graduate School of Peking Union Medical College, NO. 9, Dongdansantiao, Dongcheng District, Beijing, 100730, China.
BMC Cancer. 2019 Apr 29;19(1):405. doi: 10.1186/s12885-019-5609-1.
Wilms' tumor is also called nephroblastoma and is the most common pediatric renal cancer. Several genetic and epigenetic factors have been found to account for the development of Wilms' tumor. MiRNAs play important roles in this tumorigenic process. In the present study, we aimed to investigate the role of miR-140-5p in nephroblastoma by identifying its targets, as well as its underlying molecular mechanism of action.
The miRNA expression profile of nephroblastoma samples was investigated and the targets of miR-140-5p were predicted and validated using the miRNA luciferase reporter method. Moreover, the roles of miR-140-5p in regulating nephroblastoma cell proliferation, migration and cell cycle were analyzed by the CCK8, migration and flow cytometry assays, respectively. The downstream protein of the direct target of miR-140-5p was also identified.
miR-140-5p was downregulated in Wilms' tumor tissues, whereas in the nephroblastoma cell lines G401 and WT-CLS1 that exhibited high levels of miRNA-140-5p, inhibition of cellular proliferation and metastasis were noted as well as cell cycle arrest at the G1/S phase. TGFBRI and IGF1R were identified as direct target genes for miRNA-140-5p. In addition, SMAD2/3 and p-AKT were regulated by TGFBRI and IGF1R separately and participated in the miRNA-140-5p regulatory network. Ectopic expression of TGFBR1 and IGF-1R could abrogate the inhibitory effect of miR-140-5p.
We demonstrated that miRNA-140-5p participates in the progression of Wilms' tumor by targeting the TGFBRI/SMAD2/3 and the IGF-1R/AKT signaling pathways.
Wilms 瘤又称肾母细胞瘤，是最常见的儿童肾恶性肿瘤。已经发现几种遗传和表观遗传因素可导致 Wilms 瘤的发生。miRNA 在这一肿瘤发生过程中发挥重要作用。本研究旨在通过鉴定其靶标，以及其潜在的作用分子机制，来研究 miR-140-5p 在肾母细胞瘤中的作用。
通过 miRNA 荧光素酶报告基因法，研究了肾母细胞瘤样本的 miRNA 表达谱，并预测和验证了 miR-140-5p 的靶标。此外，通过 CCK8、迁移和流式细胞术分别分析了 miR-140-5p 对肾母细胞瘤细胞增殖、迁移和细胞周期的调控作用。还鉴定了 miR-140-5p 的直接靶标的下游蛋白。
miR-140-5p 在 Wilms 瘤组织中下调，而在 miRNA-140-5p 水平较高的肾母细胞瘤细胞系 G401 和 WT-CLS1 中，观察到细胞增殖和转移受到抑制，以及细胞周期在 G1/S 期停滞。TGFBRI 和 IGF1R 被鉴定为 miR-140-5p 的直接靶基因。此外，TGFBRI 和 IGF1R 分别调节 SMAD2/3 和 p-AKT，参与了 miR-140-5p 调控网络。TGFBR1 和 IGF-1R 的异位表达可以消除 miR-140-5p 的抑制作用。
我们证明，miR-140-5p 通过靶向 TGFBRI/SMAD2/3 和 IGF-1R/AKT 信号通路参与 Wilms 瘤的进展。
