Down-regulating NEAT1 inhibited the viability and vasculogenic mimicry formation of sinonasal squamous cell carcinoma cells via miR-195-5p/VEGFA axis.

Honglue Lu,Fei Kang

doi:10.1042/bsr20201373

Honglue Lu, Fei Kang

Open Access

https://doi.org/10.1042/bsr20201373

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Journal: Bioscience reports	Publication Date: Nov 19, 2020
Citations: 13	License type: CC BY 4.0

Affiliation: Chengde Medical University

Abstract

The role of long non-coding RNA nuclear-enriched abundant transcript 1 (lncRNA NEAT1) in sinonasal squamous cell carcinoma (SNSCC) remained obscure. Target genes and potential binding sites of NEAT1, microRNA (miR)-195-5p and VEGFA were predicted using StarBase and TargetScan, and confirmed by dual-luciferase reporter assay. Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to detect the expressions of NEAT1, vascular endothelial growth factor A (VEGFA) and miR-195-5p. Pearson’s correlation analysis of NEAT1, miR-195-5p and VEGFA was conducted. Cell viability, apoptosis and tube formation capability were assessed by MTT assay, flow cytometry and capillary-like tube formation assay, respectively. Expressions of VEGFA and proteins related to the phosphatidylinositide 3-kinase/Protein Kinase B (PI3K/AKT) pathway were measured by Western blot. In SNSCC tissues and cells, the expressions of NEAT1 and VEGFA were up-regulated while the expression of miR-195-5p was down-regulated, and NEAT1 was negatively correlated with miR-195-5p yet positively correlated with VEGFA. Overexpressed VEGFA promoted the viability and capillary-like tube formation of SNSCC cells yet suppressed their apoptosis, while silencing VEGFA led to the opposite results. MiR-195-5p could bind to NEAT1, and down-regulating miR-195-5p reversed the effects of silencing NEAT1 on the expressions of NEAT1 and miR-195-5p, cell viability, apoptosis and capillary-like tube formation as well as PI3K/AKT pathway activation. VEGFA was the target of miR-195-5p, and overexpressed VEGFA reversed the effects of miR-195-5p. Down-regulating NEAT1 inhibited the viability and vasculogenic mimicry formation of SNSCC cells yet promoted their apoptosis via the miR-195-5p/VEGFA axis, providing a possible therapeutic target for SNSCC treatment.

Highlights

Sinonasal malignancies are rarely occurring tumors which account for less than 3% of overall head and neck cancers [1]
These results suggested that miR-195-5p could bind to nuclear-enriched abundant transcript 1 (NEAT1) and target vascular endothelial growth factor A (VEGFA)
Western blot and Quantitative real-time polymerase chain reaction (qRT-PCR) results in Figure 2A–D demonstrated that both NEAT1 and VEGFA expressions were up-regulated yet miR-195-5p expression was down-regulated in sinonasal squamous cell carcinoma (SNSCC) tissues (Tumor) as compared with healthy tissues (Normal) (P

Summary

Introduction

Sinonasal malignancies are rarely occurring tumors which account for less than 3% of overall head and neck cancers [1]. Squamous cell carcinoma is the most common type of sinonasal malignancy, occupying up to 75% of all sinonasal malignancies [2]. Sinonasal malignancies are categorized as aggressive tumors because patients, who remain asymptomatic before diagnosis, are generally diagnosed at an advanced stage when the tumor has grown big enough to manifest symptoms. Though remarkable progress has been made in surgery and therapy, the prognosis of patients with sinonasal squamous cell carcinoma (SNSCC) remained poor [4]. It is of great urgency and significance to further discover the molecular mechanisms related to SNSCC development and progression

Methods

Results

Conclusion