LncRNA NEAT1 Enhances Glioma Progression via Regulating the miR-128-3p/ITGA5 Axis

Jiakai Chen,Chulei Deng,Mengliang Zhou,Handong Wang,Wenhao Niu,Junjun Wang

doi:10.1007/s12035-021-02474-y

Abstract

Accumulating evidences indicate that long non-coding RNA nuclear paraspeckle assembly transcript 1 (NEAT1) promotes the progression of glioma. In this study, we postulated that NEAT1 may act as a miR-128-3p sponge. Relative levels of NEAT1 and miR-128-3p expression in human glioma samples and GBM cells were detected using quantitative real-time PCR. By means of CCK-8 assays, transwell assays, and flow cytometric analysis, the biological functions of miR-128-3p and NEAT1 were investigated in U87MG and U251MG human GBM cell lines with stable miR-128-3p and NEAT1 knockdown or overexpression. The luciferase reports, RNA pull-down assay, and RNA immunoprecipitation assay were conducted to determine the relevance of NEAT1 and miR-128-3p in glioma. As a result, high expression of NEAT1 and lack of miR-128-3p were observed in glioma specimens and cells. By binding to anti-oncogene miR-128-3p in the nucleus, NEAT1 enhanced tumorigenesis and glioma development. Further experiments suggested that ITGA5 expression was increased in glioma tissues and was found to be connected with miR-128-3p. Additionally, NEAT1 facilitated ITGA5 expression via competitively binding to miR-128-3p. For this reason, ITGA5 would not be decomposed by miR-128-3p and could activate FAK signaling pathway, thereby promoting cell growth. Collectively, these results indicated that the NEAT1/miR-128-3p/ITGA5 axis was involved in glioma initiation and progression, and might offer a potential novel strategy for treatment of glioma.

Highlights

Among the central nervous system (CNS), glioma is the most frequently occurring primary intraparenchymal tumor [1]
nuclear paraspeckle assembly transcript 1 (NEAT1) Was Upregulated While miR-128-3p Was Downregulated in Human Glioma Tissues and Cell Lines
U87MG cells transfected with pre-miR-128-3p or pLV-Ctrl were lysed in RIPA buffer, incubated overnight with antiAGO2 antibody or IgG, followed by conjugation with protein A + G agarose to capture the antigen-antibody complex

Summary

Introduction

Among the central nervous system (CNS), glioma is the most frequently occurring primary intraparenchymal tumor [1]. Targeted molecular therapy has rapidly developed in recent years, as evidenced by a growing number of studies exploring novel promising molecular therapeutic targets for glioma [4,5,6]. LncRNA nuclear paraspeckle assembly transcript 1 (NEAT1) is an essential component of the nuclear paraspeckle substructure [9]. NEAT1 is a pro-oncogenic factor that plays a positive role in various solid tumors, such as liver, prostate, and gastric cancers, as well as renal cell carcinomas and glioma [10,11,12,13,14].

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