MiR-140-5p could suppress tumor proliferation and progression by targeting TGFBRI/SMAD2/3 and IGF-1R/AKT signaling pathways in Wilms\u2019 tumor

Zhuo Liu,Feng He,Huibo Chang,Feifei Ma,Jianxin Wu,Dingding Cao,Yuanyuan Li,Shengrong Ouyang

doi:10.1186/s12885-019-5609-1

Abstract

BackgroundWilms’ 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.MethodsThe 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.ResultsmiR-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.ConclusionWe 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.

Highlights

Wilms’ tumor is called nephroblastoma and is the most common pediatric renal cancer
MicroRNA-140-5p is downregulated in nephroblastoma tissues and is associated with clinical outcome miRCURY LNA microRNA chips from Exiqon was used to analyze three cases of nephroblastoma and adjacent normal tissue (ANT) obtained by surgical removal. miRNA microarray analysis revealed that miR-140-5p was significantly downregulated in nephroblastoma tissues compared with the corresponding levels of expression in adjacent non-cancerous tissue (ANT). quantitative Real-Time PCR (qRT-PCR) was performed in 23 cases of nephroblastoma tissues and 23 ANTs. qRT-PCR analysis revealed that the expression of miRNA-140-5p was significantly decreased in nephroblastoma tissues compared with that of the control group (Fig. 1a)
The results indicated that miRNA-140-5p may participate in Wilms’ tumor progression

Summary

Introduction

Wilms’ tumor is called nephroblastoma and is the most common pediatric renal cancer. MiRNAs play important roles in this tumorigenic process. Nephroblastoma is one of the most common solid tumours in children, with an annual incidence rate of 1 case per 100,000 children. This disease comprises 8–10% of all neoplasms in that group [1, 2]. The pathogenesis of nephroblastoma remains undiscovered, increasing evidence has suggested that multiple signalling pathways, such as microRNAs, and epigenetic mechanisms play pivotal roles in its progression. MicroRNAs (miRNAs) are endogenously produced, small (17~25-nucleotides long), non-coding single-stranded RNAs that play important roles in the regulation of crucial biological processes including cell apoptosis, metabolism, inflammation and tumorigenesis, primarily by inhibiting gene expression [3]. Yang et al demonstrated that miR-140-5p could suppress tumor progression

Objectives

Methods

Results

Discussion

Conclusion