UPF1 regulates the malignant biological behaviors of glioblastoma cells via enhancing the stability of Linc-00313

Lianqi Shao,Yixue Xue,Qianru He,Xiaobai Liu,Jun Ma,Libo Liu,Han Li,Yunhui Liu,Jian Zheng,Zhen Li

doi:10.1038/s41419-019-1845-1

Abstract

There is growing evidence that the long non-coding RNAs(lncRNAs) play an important role in the biological behaviors of glioblastoma cells. In this study, we elucidated the function and possible effect and molecular mechanisms of lncRNA-Linc-00313 on the biological behaviors of glioblastoma cells as well as UPF1 function as a RNA-binding protein to enhance its stability. Here, we used qRT-PCR and western blot to measure the expression, cell Transfection to disrupt the expression of genes, cell viability analysis, quantization of apoptosis, cell migration, and invasion assays, Reporter vectors construction and luciferase assays to investigate the malignant biological behaviors of cells, human lncRNA microarrays, RNA-Immunoprecipitation, dual-luciferase gene reporter assay, half-life assay and chromatin immunoprecipitation to verify the binding sites, tumor xenograft implantation for in vivo experiment, SPSS 18.0 statistical software for data statistics. UPF1 and Linc-00313 were both upregulated in glioma tissues and cells. Knockdown of UPF1 or Linc-00313 significantly inhibited malignant biological behaviors of glioma cells by regulating miR-342-3p and miR-485-5p, which are downregulated and functioned as tumor suppressors in glioma. Furthermore, Linc-00313 could acted as a competing endogenous RNA(ceRNA) to regulate the expression of Zic4 by binding to miR-342-3p and miR-485-5p. Interestingly, Zic4 could bind to the promoters of UPF1 and Linc-00313 respectively and upregulate the expression of them. These results indicated that a positive-feedback loop was formed in the regulation of the biological behaviors of glioma cells. The study is the first to prove that the UPF1-Linc-00313-miR-342-3p/miR-485-5p-Zic4-SHCBP1 pathway forms a positive-feedback loop and regulates the biological behaviors of U87 and U251 cells, which might provide a new therapeutic target for glioma.

Highlights

Glioblastoma is the most common primary malignant tumor of the central nervous system in adults with an annual incidence rate of approximately 6 per 100,000 persons[1]
UPF1 and Linc-00313 were upregulated in glioma tissues and cells, knockdown of UPF1, Linc-00313 inhibited the malignant biological behaviors of glioma cells
The proliferation, migration, and invasion of U87 and U251 cells were significantly decreased and the apoptosis were significantly increased in the UPF1(−) group compared with the UPF1(−)NC group (Fig. 1c–e)

Summary

Introduction

Glioblastoma is the most common primary malignant tumor of the central nervous system in adults with an annual incidence rate of approximately 6 per 100,000 persons[1]. RNA-binding proteins are important for the regulation of gene expression at a post-transcriptional level[3]. Binding proteins interact with RNA to regulate cell functions and are widely involved in RNA cleavage, transport, editing, intracellular localization, and translation regulation[4,5]. Long non-coding RNAs (LncRNAs) are a class of RNAs with transcripts longer than 200 nucleotides without protein-coding ability[6,7], RNA-. RNA-binding proteins can bind to lncRNAs and regulate its stability[10], thereby regulating the development and progression of many diseases which including tumors

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