Circ0093740 Promotes Tumor Growth and Metastasis by Sponging miR-136/145 and Upregulating DNMT3A in Wilms Tumor.

Juan Cao,Feiqiu Wen,Zhongying Huang,Sixi Liu,Xiaoxiao He,Lijuan Jiang,Huang Zhang,Yue Wang,Shunling Ou,Qiuling Miao,Yingying Shan,Guocheng Yang

doi:10.3389/fonc.2021.647352

Abstract

As a research hotspot, circular RNAs (circRNAs) is one type of non-coding RNAs which have many different functions in biological processes. However, there is lack of study investigating the underlying molecular mechanism and the potential roles of circRNAs in Wilms tumor. We conducted a high-throughput microarray sequencing to screen differentially expressed circRNAs in Wilms tumor. A novel circRNA (circ0093740) was identified as a frequently upregulated circRNA in Wilms tumor cells and tissues. Suppression of circ0093740 remarkably inhibited the proliferation and migration ability in Wilms tumor, validated by several experiments. The molecular mechanism of circ0093740 was investigated by luciferase assays and RNA immunoprecipitation assays. The results revealed that circ0093740 promotes the growth and migration ability by sponging miR-136/145 and upregulating DNMT3A. In conclusion, our study discovered the biological role of the circ0093740-miR-136/145-DNMT3A axis in Wilms tumor growth and metastasis which is important for developing new treatment strategy.

Highlights

Nephroblastoma is a kind of mixed embryonal tumor, which accounts for over 7% of all childhood malignancy and 90% of childhood renal tumors [1]
The actinomycin D assays and the RNase R assays were carried out FIGURE 1 | circ0093740 is upregulated in Wilms tumor compared to normal kidney tissue. (A) The heat map depicted the differentially expressed circRNAs of three paired Wilms tumor and normal kidney tissue. (B) The relative expression level of top five upregulated circRNA between Wilms tumor tissues and normal kidney tissues. (C) The relative expression level of circ0093740 in normal HEK293T cell line and Wilms tumor cell lines. (D) The circular structure of circ0093740 was tested RNase R assay in SKNEP1 Wilms tumor cell line. (E) Circular transcripts of circ0093740 were more stable than its linear PDCH15 mRNA transcripts determined by actinomycin D treated assay in SKNEP1 Wilms tumor cell line. **p < 0.01
Hundreds of circRNAs were discovered as novel predictive biomarkers and promising treatment targets for cancer therapies [23]. circ-CTNNB1 stimulates the Wnt signaling pathway to drive hepatocellular carcinoma progression by encoding a new 370-aa protein. circGNB1 and circRAD18 were identified as oncogenes though the mechanism of ceRNAs in breast cancer [24, 25]

Summary

INTRODUCTION

Nephroblastoma ( known as Wilms tumor) is a kind of mixed embryonal tumor, which accounts for over 7% of all childhood malignancy and 90% of childhood renal tumors [1]. CircRNAs are one novel type of single strand non-coding RNAs transcripts in cells, which play very important role in regulating genes via different molecular mechanism [4]. Taking the most famous ciRS-7/cdr1as as an example, this circRNA promotes the growth, migration, chemotherapy resistance, and immune deficiency by sponging miR-7 in multiple types of tumors [10,11,12,13,14]. Regulated by TNRC6A, circ0006916 was discovered as tumor promotor in lung cancer cells [17]. We identified a novel circRNA (circ0093740) as a frequently upregulated circRNA in Wilms tumor cells and tissues. We discovered the biological functions of the circ0093740-miR-136/ 145-DNMT3A axis in Wilms tumor growth and metastasis which is important for developing new treatment strategy

MATERIALS AND METHODS

RESULTS

C D circ0093740 in Wilms Tumor B

Findings

ETHICS STATEMENT